T cells specific for α-myosin drive immunotherapy-related myocarditis

Abstract: Immune-related adverse events, particularly severe toxicities such as myocarditis, are major challenges to the utility of immune checkpoint inhibitors (ICIs) in anticancer therapy1. The pathogenesis of ICI-associated myocarditis (ICI-MC) is poorly understood. Pdcd1-/-Ctla4+/- mice recapitulate clinicopathological features of ICI-MC, including myocardial T cell infiltration2. Here, using single-cell RNA and T cell receptor (TCR) sequencing of cardiac immune infiltrates from Pdcd1-/-Ctla4+/- mice, we identify clonal effector CD8+ T cells as the dominant cell population. Treatment with anti-CD8-depleting, but not anti-CD4-depleting, antibodies improved the survival of Pdcd1-/-Ctla4+/- mice. Adoptive transfer of immune cells from mice with myocarditis induced fatal myocarditis in recipients, which required CD8+ T cells. The cardiac-specific protein α-myosin, which is absent from the thymus3,4, was identified as the cognate antigen source for three major histocompatibility complex class I-restricted TCRs derived from mice with fulminant myocarditis. Peripheral blood T cells from three patients with ICI-MC were expanded by α-myosin peptides. Moreover, these α-myosin-expanded T cells shared TCR clonotypes with diseased heart and skeletal muscle, which indicates that α-myosin may be a clinically important autoantigen in ICI-MC. These studies underscore the crucial role for cytotoxic CD8+ T cells, identify a candidate autoantigen in ICI-MC and yield new insights into the pathogenesis of ICI toxicity.

Axelrod ML
Meijers WC
Screever EM
Qin J
Carroll MG
Sun X
Tannous E
Zhang Y
Sugiura A
Taylor BC
Hanna A
Zhang S
Amancherla K
Tai W
Wright JJ
Wei SC
Opalenik SR
Toren AL
Rathmell JC
Ferrell PB
Phillips EJ
Mallal S
Johnson DB
Allison JP
Moslehi JJ
Balko JM

Animals
Mice
Autoantigens
CD8-Positive T-Lymphocytes
CTLA-4 Antigen
Immunotherapy
Myocarditis
Ventricular Myosins
Programmed Cell Death 1 Receptor

Journal Article
Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural


Intestinal toxicity to CTLA-4 blockade driven by IL-6 and myeloid infiltration

Abstract: Immune checkpoint blockade (ICB) has revolutionized cancer treatment, yet quality of life and continuation of therapy can be constrained by immune-related adverse events (irAEs). Limited understanding of irAE mechanisms hampers development of approaches to mitigate their damage. To address this, we examined whether mice gained sensitivity to anti-CTLA-4 (αCTLA-4)-mediated toxicity upon disruption of gut homeostatic immunity. We found αCTLA-4 drove increased inflammation and colonic tissue damage in mice with genetic predisposition to intestinal inflammation, acute gastrointestinal infection, transplantation with a dysbiotic fecal microbiome, or dextran sodium sulfate administration. We identified an immune signature of αCTLA-4-mediated irAEs, including colonic neutrophil accumulation and systemic interleukin-6 (IL-6) release. IL-6 blockade combined with antibiotic treatment reduced intestinal damage and improved αCTLA-4 therapeutic efficacy in inflammation-prone mice. Intestinal immune signatures were validated in biopsies from patients with ICB colitis. Our work provides new preclinical models of αCTLA-4 intestinal irAEs, mechanistic insights into irAE development, and potential approaches to enhance ICB efficacy while mitigating irAEs.

Zhou Y
Medik YB
Patel B
Zamler DB
Chen S
Chapman T
Schneider S
Park EM
Babcock RL
Chrisikos TT
Kahn LM
Dyevoich AM
Pineda JE
Wong MC
Mishra AK
Cass SH
Cogdill AP
Johnson DH
Johnson SB
Wani K
Ledesma DA
Hudgens CW
Wang J
Wadud Khan MA
Peterson CB
Joon AY
Peng W
Li HS
Arora R
Tang X
Raso MG
Zhang X
Foo WC
Tetzlaff MT
Diehl GE
Clise-Dwyer K
Whitley EM
Gubin MM
Allison JP
Hwu P
Ajami NJ
Diab A
Wargo JA
Watowich SS

Mice
Animals
Interleukin-6
Quality of Life
Colitis
Immunotherapy
Inflammation

Research Support, U.S. Gov't, Non-P.H.S.


Immune checkpoint therapy: Forging ahead

Abstract: Spatiotemporal immune monitoring in clinical trials and reverse translation will help to determine optimal combination immune therapies to cure cancer.

Humans
Immunotherapy
Neoplasms
Combined Modality Therapy


Neoadjuvant relatlimab and nivolumab in resectable melanoma

Abstract: Relatlimab and nivolumab combination immunotherapy improves progression-free survival over nivolumab monotherapy in patients with unresectable advanced melanoma1. We investigated this regimen in patients with resectable clinical stage III or oligometastatic stage IV melanoma (NCT02519322). Patients received two neoadjuvant doses (nivolumab 480 mg and relatlimab 160 mg intravenously every 4 weeks) followed by surgery, and then ten doses of adjuvant combination therapy. The primary end point was pathologic complete response (pCR) rate2. The combination resulted in 57% pCR rate and 70% overall pathologic response rate among 30 patients treated. The radiographic response rate using Response Evaluation Criteria in Solid Tumors 1.1 was 57%. No grade 3-4 immune-related adverse events were observed in the neoadjuvant setting. The 1- and 2-year recurrence-free survival rate was 100% and 92% for patients with any pathologic response, compared to 88% and 55% for patients who did not have a pathologic response (P = 0.005). Increased immune cell infiltration at baseline, and decrease in M2 macrophages during treatment, were associated with pathologic response. Our results indicate that neoadjuvant relatlimab and nivolumab induces a high pCR rate. Safety during neoadjuvant therapy is favourable compared to other combination immunotherapy regimens. These data, in combination with the results of the RELATIVITY-047 trial1, provide further confirmation of the efficacy and safety of this new immunotherapy regimen.

Amaria RN
Postow M
Burton EM
Tetzlaff MT
Ross MI
Torres-Cabala C
Glitza IC
Duan F
Milton DR
Busam K
Simpson L
McQuade JL
Wong MK
Gershenwald JE
Lee JE
Goepfert RP
Keung EZ
Fisher SB
Betof-Warner A
Shoushtari AN
Callahan M
Coit D
Bartlett EK
Bello D
Momtaz P
Nicholas C
Gu A
Zhang X
Korivi BR
Patnana M
Patel SP
Diab A
Lucci A
Prieto VG
Davies MA
Allison JP
Sharma P
Wargo JA
Ariyan C
Tawbi HA

Humans
Antibodies, Monoclonal
Immune Checkpoint Inhibitors
Melanoma
Neoadjuvant Therapy
Neoplasm Staging
Nivolumab
Macrophages
Drug Therapy, Combination
Survival Rate
Antibodies, Monoclonal, Humanized

Clinical Trial
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't


Interleukin-6 blockade abrogates immunotherapy toxicity and promotes tumor immunity

Abstract: Immune checkpoint blockade (ICB) therapy frequently induces immune-related adverse events. To elucidate the underlying immunobiology, we performed a deep immune analysis of intestinal, colitis, and tumor tissue from ICB-treated patients with parallel studies in preclinical models. Expression of interleukin-6 (IL-6), neutrophil, and chemotactic markers was higher in colitis than in normal intestinal tissue; T helper 17 (Th17) cells were more prevalent in immune-related enterocolitis (irEC) than T helper 1 (Th1). Anti-cytotoxic T-lymphocyte-associated antigen 4 (anti-CTLA-4) induced stronger Th17 memory in colitis than anti-program death 1 (anti-PD-1). In murine models, IL-6 blockade associated with improved tumor control and a higher density of CD4+/CD8+ effector T cells, with reduced Th17, macrophages, and myeloid cells. In an experimental autoimmune encephalomyelitis (EAE) model with tumors, combined IL-6 blockade and ICB enhanced tumor rejection while simultaneously mitigating EAE symptoms versus ICB alone. IL-6 blockade with ICB could de-couple autoimmunity from antitumor immunity.

Hailemichael Y
Johnson DH
Abdel-Wahab N
Foo WC
Bentebibel SE
Daher M
Haymaker C
Wani K
Saberian C
Ogata D
Kim ST
Nurieva R
Lazar AJ
Abu-Sbeih H
Fa'ak F
Mathew A
Wang Y
Falohun A
Trinh V
Zobniw C
Spillson C
Burks JK
Awiwi M
Elsayes K
Soto LS
Melendez BD
Davies MA
Wargo J
Curry J
Yee C
Lizee G
Singh S
Sharma P
Allison JP
Hwu P
Ekmekcioglu S
Diab A

Animals
Colitis
Humans
Immunologic Factors
Immunotherapy
Interleukin-6
Mice
Myeloid Cells
Neoplasms


A phase 1-2 trial of sitravatinib and nivolumab in clear cell renal cell carcinoma following progression on antiangiogenic therapy

Abstract: The accumulation of immune-suppressive myeloid cells is a critical determinant of resistance to anti-programmed death-1 (PD-1) therapy in advanced clear cell renal cell carcinoma (ccRCC). In preclinical models, the tyrosine kinase inhibitor sitravatinib enhanced responses to anti-PD-1 therapy by modulating immune-suppressive myeloid cells. We conducted a phase 1-2 trial to choose an optimal sitravatinib dose combined with a fixed dose of nivolumab in 42 immunotherapy-naïve patients with ccRCC refractory to prior antiangiogenic therapies. The combination demonstrated no unexpected toxicities and achieved an objective response rate of 35.7% and a median progression-free survival of 11.7 months, with 80.1% of patients alive after a median follow-up of 18.7 months. Baseline peripheral blood neutrophil-to-lymphocyte ratio correlated with response to sitravatinib and nivolumab. Patients with liver metastases showed durable responses comparable to patients without liver metastases. In addition, correlative studies demonstrated reduction of immune-suppressive myeloid cells in the periphery and tumor microenvironment following sitravatinib treatment. This study provides a rationally designed combinatorial strategy to improve outcomes of anti-PD-1 therapy in advanced ccRCC.

Msaouel P
Goswami S
Thall PF
Wang X
Yuan Y
Jonasch E
Gao J
Campbell MT
Shah AY
Corn PG
Tam AL
Ahrar K
Rao P
Sircar K
Cohen L
Basu S
Duan F
Jindal S
Zhang Y
Chen H
Yadav SS
Shazer R
Der-Torossian H
Allison JP
Sharma P
Tannir NM

Angiogenesis Inhibitors
Anilides
Carcinoma, Renal Cell
Female
Humans
Kidney Neoplasms
Liver Neoplasms
Male
Nivolumab
Pyridines
Tumor Microenvironment

Clinical Trial, Phase I
Clinical Trial, Phase II
Journal Article
Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural


Correction: Pilot Phase II Trial of Neoadjuvant Immunotherapy in Locoregionally Advanced, Resectable Cutaneous Squamous Cell Carcinoma of the Head and Neck

Clinical cancer research : an official journal of the American Association for Cancer Research

Ferrarotto R
Amit M
Nagarajan P
Rubin ML
Yuan Y
Bell D
El-Naggar AK
Johnson JM
Morrison WH
Rosenthal DI
Glisson BS
Johnson FM
Lu C
Mott FE
Esmaeli B
Diaz EM
Gidley PW
Goepfert RP
Lewis CM
Weber RS
Wargo JA
Basu S
Duan F
Yadav SS
Sharma P
Allison JP
Myers JN
Gross ND


Perioperative nivolumab monotherapy versus nivolumab plus ipilimumab in resectable hepatocellular carcinoma: a randomised, open-label, phase 2 trial

Abstract: Hepatocellular carcinoma has high recurrence rates after surgery; however, there are no approved standard-of-care neoadjuvant or adjuvant therapies. Immunotherapy has been shown to improve survival in advanced hepatocellular carcinoma; we therefore aimed to evaluate the safety and tolerability of perioperative immunotherapy in resectable hepatocellular carcinoma. In this single-centre, randomised, open-label, phase 2 trial, patients with resectable hepatocellular carcinoma were randomly assigned (1:1) to receive 240 mg of nivolumab intravenously every 2 weeks (for up to three doses before surgery at 6 weeks) followed in the adjuvant phase by 480 mg of nivolumab intravenously every 4 weeks for 2 years, or 240 mg of nivolumab intravenously every 2 weeks (for up to three doses before surgery) plus one dose of 1 mg/kg of ipilimumab intravenously concurrently with the first preoperative dose of nivolumab, followed in the adjuvant phase by 480 mg of nivolumab intravenously every 4 weeks for up to 2 years plus 1 mg/kg of ipilimumab intravenously every 6 weeks for up to four cycles. Patients were randomly assigned to the treatment groups by use of block randomisation with a random block size. The primary endpoint was the safety and tolerability of nivolumab with or without ipilimumab. Secondary endpoints were the proportion of patients with an overall response, time to progression, and progression-free survival. This trial is registered with ClinicalTrials.gov (NCT03222076) and is completed. Between Oct 30, 2017, and Dec 3, 2019, 30 patients were enrolled and 27 were randomly assigned: 13 to nivolumab and 14 to nivolumab plus ipilimumab. Grade 3-4 adverse events were higher with nivolumab plus ipilimumab (six [43%] of 14 patients) than with nivolumab alone (three [23%] of 13). The most common treatment-related adverse events of any grade were increased alanine aminotransferase (three [23%] of 13 patients on nivolumab vs seven [50%] of 14 patients on nivolumab plus ipilimumab) and increased aspartate aminotransferase (three [23%] vs seven [50%]). No patients in either group had their surgery delayed due to grade 3 or worse adverse events. Seven of 27 patients had surgical cancellations, but none was due to treatment-related adverse events. Estimated median progression-free survival was 9·4 months (95% CI 1·47-not estimable [NE]) with nivolumab and 19·53 months (2·33-NE) with nivolumab plus ipilimumab (hazard ratio [HR] 0·99, 95% CI 0·31-2·54); median time to progression was 9·4 months (95% CI 1·47-NE) in the nivolumab group and 19·53 months (2·33-NE) in the nivolumab plus ipilimumab group (HR 0·89, 95% CI 0·31-2·54). In an exploratory analysis, three (23%) of 13 patients had an overall response with nivolumab monotherapy, versus none with nivolumab plus ipilimumab. Three (33%) of nine patients had a major pathological response (ie, ≥70% necrosis in the resected tumour area) with nivolumab monotherapy compared with three (27%) of 11 with nivolumab plus ipilimumab. Perioperative nivolumab alone and nivolumab plus ipilimumab appears to be safe and feasible in patients with resectable hepatocellular carcinoma. Our findings support further studies of immunotherapy in the perioperative setting in hepatocellular carcinoma. Bristol Myers Squibb and the US National Institutes of Health.

The lancet. Gastroenterology & hepatology

Kaseb AO
Hasanov E
Cao HST
Xiao L
Vauthey JN
Lee SS
Yavuz BG
Mohamed YI
Qayyum A
Jindal S
Duan F
Basu S
Yadav SS
Nicholas C
Sun JJ
Singh Raghav KP
Rashid A
Carter K
Chun YS
Tzeng CD
Sakamuri D
Xu L
Sun R
Cristini V
Beretta L
Yao JC
Wolff RA
Allison JP
Sharma P

Antineoplastic Combined Chemotherapy Protocols

Clinical Trial, Phase II
Journal Article
Randomized Controlled Trial
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't


Checkpoint Blockade + Chemotherapy: the Right Combination for AML?

Abstract: An emerging strategy to enhance the efficacy of immune checkpoint blockade in relapsed/refractory cancers is increasing immunogenic cell death via combination with cytotoxic therapies. Understanding the effects of cytotoxic and immunotherapeutic agents on immune cell populations will enable improved mechanism-based design of combination therapies to maximum efficacy and minimum toxicity.See related article by Zeidner et al., p. 616.


Dietary fiber and probiotics influence the gut microbiome and melanoma immunotherapy response

Abstract: Gut bacteria modulate the response to immune checkpoint blockade (ICB) treatment in cancer, but the effect of diet and supplements on this interaction is not well studied. We assessed fecal microbiota profiles, dietary habits, and commercially available probiotic supplement use in melanoma patients and performed parallel preclinical studies. Higher dietary fiber was associated with significantly improved progression-free survival in 128 patients on ICB, with the most pronounced benefit observed in patients with sufficient dietary fiber intake and no probiotic use. Findings were recapitulated in preclinical models, which demonstrated impaired treatment response to anti–programmed cell death 1 (anti–PD-1)–based therapy in mice receiving a low-fiber diet or probiotics, with a lower frequency of interferon-γ–positive cytotoxic T cells in the tumor microenvironment. Together, these data have clinical implications for patients receiving ICB for cancer.

Spencer CN
McQuade JL
Gopalakrishnan V
McCulloch JA
Vetizou M
Cogdill AP
Khan MAW
Zhang X
White MG
Peterson CB
Wong MC
Morad G
Rodgers T
Badger JH
Helmink BA
Andrews MC
Rodrigues RR
Morgun A
Kim YS
Roszik J
Hoffman KL
Zheng J
Zhou Y
Medik YB
Kahn LM
Johnson S
Hudgens CW
Wani K
Gaudreau PO
Harris AL
Jamal MA
Baruch EN
Perez-Guijarro E
Day CP
Merlino G
Pazdrak B
Lochmann BS
Szczepaniak-Sloane RA
Arora R
Anderson J
Zobniw CM
Posada E
Sirmans E
Simon J
Haydu LE
Burton EM
Wang L
Dang M
Clise-Dwyer K
Schneider S
Chapman T
Anang NAS
Duncan S
Toker J
Malke JC
Glitza IC
Amaria RN
Tawbi HA
Diab A
Wong MK
Patel SP
Woodman SE
Davies MA
Ross MI
Gershenwald JE
Lee JE
Hwu P
Jensen V
Samuels Y
Straussman R
Ajami NJ
Nelson KC
Nezi L
Petrosino JF
Futreal PA
Lazar AJ
Hu J
Jenq RR
Tetzlaff MT
Yan Y
Garrett WS
Huttenhower C
Sharma P
Watowich SS
Allison JP
Cohen L
Trinchieri G
Daniel CR
Wargo JA

Animals
Cohort Studies
Dietary Fiber
Fatty Acids, Volatile
Fecal Microbiota Transplantation
Feces
Female
Gastrointestinal Microbiome
Humans
Immune Checkpoint Inhibitors
Immunotherapy
Male
Melanoma
Melanoma, Experimental
Mice
Mice, Inbred C57BL
Probiotics
Progression-Free Survival
T-Lymphocytes

Journal Article
Observational Study
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't


Pilot study of Tremelimumab with and without cryoablation in patients with metastatic renal cell carcinoma

Abstract: Cryoablation in combination with immune checkpoint therapy was previously reported to improve anti-tumor immune responses in pre-clinical studies. Here we report a pilot study of anti-CTLA-4 (tremelimumab) with (n = 15) or without (n = 14) cryoablation in patients with metastatic renal cell carcinoma (NCT02626130), 18 patients with clear cell and 11 patients with non-clear cell histologies. The primary endpoint is safety, secondary endpoints include objective response rate, progression-free survival, and immune monitoring studies. Safety data indicate ≥ grade 3 treatment-related adverse events in 16 of 29 patients (55%) including 6 diarrhea/colitis, 3 hepatitis, 1 pneumonitis, and 1 glomerulonephritis. Toxicity leading to treatment discontinuation occurs in 5 patients in each arm. 3 patients with clear cell histology experience durable responses. One patient in the tremelimumab arm experiences an objective response, the median progression-free survival for all patients is 3.3 months (95% CI: 2.0, 5.3 months). Exploratory immune monitoring analysis of baseline and post-treatment tumor tissue samples shows that treatment increases immune cell infiltration and tertiary lymphoid structures in clear cell but not in non-clear cell. In clear cell, cryoablation plus tremelimumab leads to a significant increase in immune cell infiltration. These data highlight that treatment with tremelimumab plus cryotherapy is feasible and modulates the immune microenvironment in patients with metastatic clear cell histology.

Campbell MT
Matin SF
Tam AL
Sheth RA
Ahrar K
Tidwell RS
Rao P
Karam JA
Wood CG
Tannir NM
Jonasch E
Gao J
Zurita AJ
Shah AY
Jindal S
Duan F
Basu S
Chen H
Espejo AB
Allison JP
Yadav SS
Sharma P

Adult
Aged
Aged, 80 and over
Antibodies, Monoclonal, Humanized
Antineoplastic Agents, Immunological
CTLA-4 Antigen
Carcinoma, Renal Cell
Combined Modality Therapy
Cryosurgery
Female
Humans
Kidney Neoplasms
Male
Middle Aged
Neoplasm Metastasis
Patient Safety
Pilot Projects
Survival Rate
Treatment Outcome
Young Adult

Clinical Trial, Phase I
Journal Article
Randomized Controlled Trial
Research Support, Non-U.S. Gov't


Single cell T cell landscape and T cell receptor repertoire profiling of AML in context of PD-1 blockade therapy

Abstract: In contrast to the curative effect of allogenic stem cell transplantation in acute myeloid leukemia via T cell activity, only modest responses are achieved with checkpoint-blockade therapy, which might be explained by T cell phenotypes and T cell receptor (TCR) repertoires. Here, we show by paired single-cell RNA analysis and TCR repertoire profiling of bone marrow cells in relapsed/refractory acute myeloid leukemia patients pre/post azacytidine+nivolumab treatment that the disease-related T cell subsets are highly heterogeneous, and their abundance changes following PD-1 blockade-based treatment. TCR repertoires expand and primarily emerge from CD8+ cells in patients responding to treatment or having a stable disease, while TCR repertoires contract in therapy-resistant patients. Trajectory analysis reveals a continuum of CD8+ T cell phenotypes, characterized by differential expression of granzyme B and a bone marrow-residing memory CD8+ T cell subset, in which a population with stem-like properties expressing granzyme K is enriched in responders. Chromosome 7/7q loss, on the other hand, is a cancer-intrinsic genomic marker of PD-1 blockade resistance in AML. In summary, our study reveals that adaptive T cell plasticity and genomic alterations determine responses to PD-1 blockade in acute myeloid leukemia.

Abbas HA
Hao D
Tomczak K
Barrodia P
Im JS
Reville PK
Alaniz Z
Wang W
Wang R
Wang F
Al-Atrash G
Takahashi K
Ning J
Ding M
Beird HC
Mathews JT
Little L
Zhang J
Basu S
Konopleva M
Marques-Piubelli ML
Solis LM
Parra ER
Lu W
Tamegnon A
Garcia-Manero G
Green MR
Sharma P
Allison JP
Kornblau SM
Rai K
Wang L
Daver N
Futreal A

Aged
Aged, 80 and over
Azacitidine
Bone Marrow
CD8-Positive T-Lymphocytes
Chromosome Deletion
Chromosomes, Human, Pair 7
Drug Resistance, Neoplasm
Granzymes
Humans
Immune Checkpoint Inhibitors
Leukemia, Myeloid, Acute
Middle Aged
Nivolumab
Programmed Cell Death 1 Receptor
Receptors, Antigen, T-Cell
Single-Cell Analysis
T-Lymphocyte Subsets
T-Lymphocytes
Transcriptome

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't


Combined CTLA-4 and PD-L1 blockade in patients with chemotherapy-naïve metastatic castration-resistant prostate cancer is associated with increased myeloid and neutrophil immune subsets in the bone microenvironment

Abstract: Immune checkpoint therapy (ICT) has low response rates in patients with metastatic castration-resistant prostate cancer (mCRPC), in part due to few T cells in the tumor microenvironment (TME). Anti-cytotoxic T lymphocyte-associated protein 4 (CTLA-4) promotes intratumoral T cell infiltration but induces upregulation of PD-1 and programmed death ligand-1 (PD-L1) within the prostate TME. Combined anti-CTLA-4 plus anti-PD-1 can partly overcome this adaptive resistance and was recently shown to augment responses in patients with mCRPC with measurable disease. Although bone is the most common site of metastasis in prostate cancer, patients with bone-predominant disease are frequently excluded from trials because they lack measurable disease, which limits assessment of disease progression and tissue sampling. We therefore designed this study to investigate combined ICT in mCRPC to bone. Combined anti-CTLA-4 (tremelimumab) plus anti-PD-L1 (durvalumab) is safe and well tolerated in patients with chemotherapy-naïve mCRPC to bone. In this single-arm pilot study, men with chemotherapy-naïve mCRPC to bone received tremelimumab (75 mg intravenous) plus durvalumab (1500 mg intravenous) every 4 weeks (up to four doses), followed by durvalumab (1500 mg intravenous) maintenance every 4 weeks (up to nine doses). The primary endpoint was incidence of adverse events. Secondary endpoints included serum prostate-specific antigen (PSA), progression-free survival (PFS), radiographic PFS (rPFS), and maximal PSA decline. Twenty-six patients were treated between August 8, 2017 and March 28, 2019. Grade ≥3 treatment-related adverse events (TRAEs) occurred in 11 patients (42%), with no grade 4 or 5 events. TRAEs leading to discontinuation occurred in three patients (12%). PSA decline ≥50% occurred in three patients (12%). Six patients (24%) achieved stable disease for >6 months. At a median follow-up of 43.6 months, median rPFS was 3.7 months (95% CI: 1.9 to 5.7), and median overall survival was 28.1 months (95% CI: 14.5 to 37.3). Post-treatment evaluation of the bone microenvironment revealed transcriptional upregulation in myeloid and neutrophil immune subset signatures and increased expression of inhibitory immune checkpoints. Tremelimumab plus durvalumab was safe and well tolerated in patients with chemotherapy-naïve mCRPC to bone, with potential activity in a small number of patients as measured by rPFS. Combination of CTLA-4 and PD-L1 blockade with therapies targeting the myeloid compartment or other inhibitory immune receptors may be necessary to overcome mechanisms of resistance within prostate bone microenvironment. NCT03204812.

Subudhi SK
Siddiqui BA
Aparicio AM
Yadav SS
Basu S
Chen H
Jindal S
Tidwell RSS
Varma A
Logothetis CJ
Allison JP
Corn PG
Sharma P


Nodal immune flare mimics nodal disease progression following neoadjuvant immune checkpoint inhibitors in non-small cell lung cancer

Abstract: Radiographic imaging is the standard approach for evaluating the disease involvement of lymph nodes in patients with operable NSCLC although the impact of neoadjuvant immune checkpoint inhibitors (ICIs) on lymph nodes has not yet been characterized. Herein, we present an ad hoc analysis of the NEOSTAR trial (NCT03158129) where we observed a phenomenon we refer to as "nodal immune flare" (NIF) in which patients treated with neoadjuvant ICIs demonstrate radiologically abnormal nodes post-therapy that upon pathological evaluation are devoid of cancer and demonstrate de novo non-caseating granulomas. Abnormal lymph nodes are analyzed by computed tomography and 18F-fluorodeoxyglucose positron emission tomography/computer tomography to evaluate the size and the maximum standard uptake value post- and pre-therapy in NEOSTAR and an independent neoadjuvant chemotherapy cohort. NIF occurs in 16% (7/44) of patients treated with ICIs but in 0% (0/28) of patients after neoadjuvant chemotherapy. NIF is associated with an inflamed nodal immune microenvironment and with fecal abundance of genera belonging to the family Coriobacteriaceae of phylum Actinobacteria, but not with tumor responses or treatment-related toxicity. Our findings suggest that this apparent radiological cancer progression in lymph nodes may occur due to an inflammatory response after neoadjuvant immunotherapy, and such cases should be evaluated by pathological examination to distinguish NIF from true nodal progression and to ensure appropriate clinical treatment planning.

Cascone T
Weissferdt A
Godoy MCB
William WN Jr
Leung CH
Lin HY
Basu S
Yadav SS
Pataer A
Mitchell KG
Khan MAW
Shi Y
Haymaker C
Solis LM
Parra ER
Kadara H
Wistuba II
Sharma P
Allison JP
Ajami NJ
Wargo JA
Jenq RR
Gibbons DL
Lee JJ
Swisher SG
Vaporciyan AA
Heymach JV
Sepesi B

Aged
Carcinoma, Non-Small-Cell Lung
Disease Progression
Female
Humans
Immune Checkpoint Inhibitors
Lung Neoplasms
Lymph Nodes
Lymphatic Metastasis
Male
Middle Aged
Multimodal Imaging
Neoadjuvant Therapy


Gut microbiota signatures are associated with toxicity to combined CTLA-4 and PD-1 blockade

Abstract: Treatment with combined immune checkpoint blockade (CICB) targeting CTLA-4 and PD-1 is associated with clinical benefit across tumor types, but also a high rate of immune-related adverse events. Insights into biomarkers and mechanisms of response and toxicity to CICB are needed. To address this, we profiled the blood, tumor and gut microbiome of 77 patients with advanced melanoma treated with CICB, with a high rate of any ≥grade 3 immune-related adverse events (49%) with parallel studies in pre-clinical models. Tumor-associated immune and genomic biomarkers of response to CICB were similar to those identified for ICB monotherapy, and toxicity from CICB was associated with a more diverse peripheral T-cell repertoire. Profiling of gut microbiota demonstrated a significantly higher abundance of Bacteroides intestinalis in patients with toxicity, with upregulation of mucosal IL-1β in patient samples of colitis and in pre-clinical models. Together, these data offer potential new therapeutic angles for targeting toxicity to CICB.

Andrews MC
Duong CPM
Gopalakrishnan V
Iebba V
Chen WS
Derosa L
Khan MAW
Cogdill AP
White MG
Wong MC
Ferrere G
Fluckiger A
Roberti MP
Opolon P
Alou MT
Yonekura S
Roh W
Spencer CN
Curbelo IF
Vence L
Reuben A
Johnson S
Arora R
Morad G
Lastrapes M
Baruch EN
Little L
Gumbs C
Cooper ZA
Prieto PA
Wani K
Lazar AJ
Tetzlaff MT
Hudgens CW
Callahan MK
Adamow M
Postow MA
Ariyan CE
Gaudreau PO
Nezi L
Raoult D
Mihalcioiu C
Elkrief A
Pezo RC
Haydu LE
Simon JM
Tawbi HA
McQuade J
Hwu P
Hwu WJ
Amaria RN
Burton EM
Woodman SE
Watowich S
Diab A
Patel SP
Glitza IC
Wong MK
Zhao L
Zhang J
Ajami NJ
Petrosino J
Jenq RR
Davies MA
Gershenwald JE
Futreal PA
Sharma P
Allison JP
Routy B
Zitvogel L
Wargo JA

Animals
CTLA-4 Antigen
Cell Line, Tumor
Female
Gastrointestinal Microbiome
Humans
Interleukin-1beta
Melanoma
Mice
Mice, Inbred C57BL
Programmed Cell Death 1 Receptor


Pilot Phase II Trial of Neoadjuvant Immunotherapy in Locoregionally Advanced, Resectable Cutaneous Squamous Cell Carcinoma of the Head and Neck

Abstract: In locoregionally advanced, resectable cutaneous squamous cell carcinoma of the head and neck (CSCC-HN), surgery followed by radiotherapy is standard but can be cosmetically and functionally devastating, and many patients will have recurrence. Newly diagnosed or recurrent stage III-IVA CSCC-HN patients amenable to curative-intent surgery received two cycles of neoadjuvant PD-1 inhibition. The primary endpoint was ORR per RECIST 1.1. Secondary endpoints included pathologic response [pathologic complete response (pCR) or major pathologic response (MPR; ≤10% viable tumor)], safety, DSS, DFS, and OS. Exploratory endpoints included immune biomarkers of response. Of 20 patients enrolled, 7 had recurrent disease. While only 6 patients [30%; 95% confidence interval (CI), 11.9-54.3] had partial responses by RECIST, 14 patients (70%; 95% CI, 45.7-88.1) had a pCR (n = 11) or MPR (n = 3). No SAEs ocurred during or after the neoadjuvant treatment. At a median follow-up of 22.6 months (95% CI, 21.7-26.1), one patient progressed and died, one died without disease, and two developed recurrence. The 12-month DSS, DFS, and OS rates were 95% (95% CI, 85.9-100), 89.5% (95% CI, 76.7-100), and 95% (95% CI, 85.9-100), respectively. Gene expression studies revealed an inflamed tumor microenvironment in patients with pCR or MPR, and CyTOF analyses demonstrated a memory CD8+ T-cell cluster enriched in patients with pCR. Neoadjuvant immunotherapy in locoregionally advanced, resectable CSCC-HN is safe and induces a high pathologic response rate. Pathologic responses were associated with an inflamed tumor microenvironment.

Ferrarotto R
Amit M
Nagarajan P
Rubin ML
Yuan Y
Bell D
El-Naggar AK
Johnson JM
Morrison WH
Rosenthal DI
Glisson BS
Johnson FM
Lu C
Mott FE
Esmaeli B
Diaz EM Jr
Gidley PW
Goepfert RP
Lewis CM
Weber RS
Wargo JA
Basu S
Duan F
Yadav SS
Sharma P
Allison JP
Myers JN
Gross ND

Squamous Cell Carcinoma of Head and Neck

Clinical Trial, Phase II
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't


Pre-existing immune status associated with response to combination of sipuleucel-T and ipilimumab in patients with metastatic castration-resistant prostate cancer

Abstract: Sipuleucel-T is a US Food and Drug Administration-approved autologous cellular immunotherapy that improves survival in patients with metastatic castration-resistant prostate cancer (mCRPC). We examined whether administering ipilimumab after sipuleucel-T could modify immune and/or clinical responses to this treatment. A total of 50 patients with mCRPC were enrolled into a clinical trial (NCT01804465, ClinicalTrials.gov) where they received ipilimumab either immediately or delayed 3 weeks following completion of sipuleucel-T treatment. Blood was collected at various timepoints of the study. Luminex assay for anti-prostatic acid phosphatase (PAP) and anti-PA2024-specific serum immunoglobulin G (IgG) and ELISpot for interferon-γ (IFN-γ) production against PAP and PA2024 were used to assess antigen-specific B and T cell responses, respectively. Clinical response was defined as >30% reduction in serum prostate-specific antigen levels compared with pretreatment levels. The frequency and state of circulating immune cells were determined by mass cytometry by time-of-flight and statistical scaffold analysis. We found the combination to be well tolerated with no unexpected adverse events occurring. The timing of ipilimumab did not significantly alter the rates of antigen-specific B and T cell responses, the primary endpoint of the clinical trial. Clinical responses were observed in 6 of 50 patients, with 3 having responses lasting longer than 3 months. The timing of ipilimumab did not significantly associate with clinical response or toxicity. The combination treatment did induce CD4 and CD8 T cell activation that was most pronounced with the immediate schedule. Lower frequencies of CTLA-4 positive circulating T cells, even prior to treatment, were associated with better clinical outcomes. Interestingly, these differences in CTLA-4 expression were associated with prior localized radiation therapy (RT) to the prostate or prostatic fossa. Prior radiation treatment was also associated with improved radiographic progression-free survival. Combining CTLA-4 blockade with sipuleucel-T resulted in modest clinical activity. The timing of CTLA-4 blockade following sipuleucel-T did not alter antigen-specific responses. Clinical responses were associated with both lower baseline frequencies of CTLA-4 expressing T cells and a history of RT. Prior cancer therapy may therefore result in long-lasting immune changes that influence responsiveness to immunotherapy with sipuleucel-T and anti-CTLA-4.

Sinha M
Zhang L
Subudhi S
Chen B
Marquez J
Liu EV
Allaire K
Cheung A
Ng S
Nguyen C
Friedlander TW
Aggarwal R
Spitzer M
Allison JP
Small EJ
Sharma P
Fong L

Clinical Trial, Phase II
Journal Article
Multicenter Study
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't


LILRB4 suppresses immunity in solid tumors and is a potential target for immunotherapy

Abstract: Immune receptors expressed on TAMs are intriguing targets for tumor immunotherapy. In this study, we found inhibitory receptor LILRB4 on a variety of intratumoral immune cell types in murine tumor models and human cancers, most prominently on TAMs. LILRB4, known as gp49B in mice, is a LILRB family receptor. Human and murine LILRB4 have two extracellular domains but differ in the number of intracellular ITIMs (three versus two). We observed a high correlation in LILRB4 expression with other immune inhibitory receptors. After tumor challenge, LILRB4-/- mice and mice treated with anti-LILRB4 antibody showed reduced tumor burden and increased survival. LILRB4-/- genotype or LILRB4 blockade increased tumor immune infiltrates and the effector (Teff) to regulatory (Treg) T cell ratio and modulated phenotypes of TAMs toward less suppressive, CD4+ T cells to Th1 effector, and CD8+ T cells to less exhausted. These findings reveal that LILRB4 strongly suppresses tumor immunity in TME and that alleviating that suppression provides antitumor efficacy.

Animals
CD4-Positive T-Lymphocytes
CD8-Positive T-Lymphocytes
Cell Line, Tumor
Cell Movement
Humans
Immunotherapy
Membrane Glycoproteins
Mice
Mice, Inbred C57BL
Neoplasms
Receptors, Immunologic
T-Lymphocytes, Regulatory

Journal Article
Research Support, Non-U.S. Gov't


First-in-Human Phase I Study of ABBV-085, an Antibody-Drug Conjugate Targeting LRRC15, in Sarcomas and Other Advanced Solid Tumors

Abstract: Leucine-rich repeat containing 15 (LRRC15) is expressed on stromal fibroblasts in the tumor microenvironment of multiple solid tumor types and may represent an interesting target for therapy, particularly in patients with sarcomas where LRRC15 is also expressed by malignant cells. ABBV-085 is a monomethyl auristatin-E antibody-drug conjugate that targets LRRC15 and showed antineoplastic efficacy in preclinical experiments. Herein, we report findings of ABBV-085 monotherapy or combination therapy in adult patients with sarcomas and other advanced solid tumors. This first-in-human phase I study (NCT02565758) assessed ABBV-085 safety, pharmacokinetics/pharmacodynamics, and preliminary antitumor activity. The study consisted of two parts: dose escalation and dose expansion. ABBV-085 was administered by intravenous infusion at 0.3 to 6.0 mg/kg every 14 days. In total, 85 patients were enrolled; 45 patients received the recommended expansion dose of 3.6 mg/kg ABBV-085 monotherapy, including 10 with osteosarcoma and 10 with undifferentiated pleomorphic sarcoma (UPS). Most common treatment-related adverse events were fatigue, nausea, and decreased appetite. The overall response rate for patients with osteosarcoma/UPS treated at 3.6 mg/kg was 20%, including four confirmed partial responses. No monotherapy responses were observed for other advanced cancers treated at 3.6 mg/kg. One patient treated with ABBV-085 plus gemcitabine achieved partial response. ABBV-085 appeared safe and tolerable at a dose of 3.6 mg/kg every 14 days, with preliminary antitumor activity noted in patients with osteosarcoma and UPS. Given the high unmet need in these orphan malignancies, further investigation into targeting LRRC15 in these sarcomas may be warranted.

Demetri GD
Luke JJ
Hollebecque A
Powderly JD 2nd
Spira AI
Subbiah V
Naumovski L
Chen C
Fang H
Lai DW
Yue H
Polepally AR
Purcell JW
Robinson R
Sharma P
Allison JP
Tolcher A
Villalobos VM

Adult
Antineoplastic Agents
Bone Neoplasms
Humans
Immunoconjugates
Membrane Proteins
Neoplasms
Sarcoma
Tumor Microenvironment


The Next Decade of Immune Checkpoint Therapy

Abstract: Immune checkpoint therapy (ICT) can provide durable clinical responses and improve overall survival. However, only subsets of patients with specific tumor types respond to ICT. Thus, significant challenges remain, including understanding pathways of resistance, optimizing patient selection, improving management of immune-related adverse events, and identifying rational therapeutic combinations. These challenges will need a focused approach encompassing both clinical and basic research, with the integration of reverse translational studies. This integrated approach will lead to identification of potential targets for subsequent clinical trials, which will guide decisions as we develop novel combination strategies to maximize efficacy and minimize toxicities for patients. SIGNIFICANCE: ICTs induce durable antitumor responses for subsets of patients with cancer. Recent evidence suggests that rational combinatorial strategies can improve response by overcoming primary and adaptive resistance mechanisms, although these may carry an increased risk of immune-mediated toxicities. This review surveys the current understanding of mechanisms of response and resistance to ICTs and active areas of investigation, and proposes a path forward to improving efficacy and minimizing toxicities through better patient selection and rational combinations.

Sharma P
Siddiqui BA
Anandhan S
Yadav SS
Subudhi SK
Gao J
Goswami S
Allison JP

Drug Discovery
Forecasting
Humans
Immune Checkpoint Inhibitors
Neoplasms


Neoadjuvant nivolumab or nivolumab plus ipilimumab in operable non-small cell lung cancer: the phase 2 randomized NEOSTAR trial

Abstract: Ipilimumab improves clinical outcomes when combined with nivolumab in metastatic non-small cell lung cancer (NSCLC), but its efficacy and impact on the immune microenvironment in operable NSCLC remain unclear. We report the results of the phase 2 randomized NEOSTAR trial (NCT03158129) of neoadjuvant nivolumab or nivolumab + ipilimumab followed by surgery in 44 patients with operable NSCLC, using major pathologic response (MPR) as the primary endpoint. The MPR rate for each treatment arm was tested against historical controls of neoadjuvant chemotherapy. The nivolumab + ipilimumab arm met the prespecified primary endpoint threshold of 6 MPRs in 21 patients, achieving a 38% MPR rate (8/21). We observed a 22% MPR rate (5/23) in the nivolumab arm. In 37 patients resected on trial, nivolumab and nivolumab + ipilimumab produced MPR rates of 24% (5/21) and 50% (8/16), respectively. Compared with nivolumab, nivolumab + ipilimumab resulted in higher pathologic complete response rates (10% versus 38%), less viable tumor (median 50% versus 9%), and greater frequencies of effector, tissue-resident memory and effector memory T cells. Increased abundance of gut Ruminococcus and Akkermansia spp. was associated with MPR to dual therapy. Our data indicate that neoadjuvant nivolumab + ipilimumab-based therapy enhances pathologic responses, tumor immune infiltrates and immunologic memory, and merits further investigation in operable NSCLC.

Cascone T
William WN Jr
Weissferdt A
Leung CH
Lin HY
Pataer A
Godoy MCB
Carter BW
Federico L
Reuben A
Khan MAW
Dejima H
Francisco-Cruz A
Parra ER
Solis LM
Fujimoto J
Tran HT
Kalhor N
Fossella FV
Mott FE
Tsao AS
Blumenschein G Jr
Le X
Zhang J
Skoulidis F
Kurie JM
Altan M
Lu C
Glisson BS
Byers LA
Elamin YY
Mehran RJ
Rice DC
Walsh GL
Hofstetter WL
Roth JA
Antonoff MB
Kadara H
Haymaker C
Bernatchez C
Ajami NJ
Jenq RR
Sharma P
Allison JP
Futreal A
Wargo JA
Wistuba II
Swisher SG
Lee JJ
Gibbons DL
Vaporciyan AA
Heymach JV
Sepesi B


A Genetic Mouse Model Recapitulates Immune Checkpoint Inhibitor-Associated Myocarditis and Supports a Mechanism-Based Therapeutic Intervention

Abstract: Immune checkpoint inhibitors (ICI) targeting CTLA4 or PD-1/PD-L1 have transformed cancer therapy but are associated with immune-related adverse events, including myocarditis. Here, we report a robust preclinical mouse model of ICI-associated myocarditis in which monoallelic loss of Ctla4 in the context of complete genetic absence of Pdcd1 leads to premature death in approximately half of mice. Premature death results from myocardial infiltration by T cells and macrophages and severe ECG abnormalities, closely recapitulating the clinical and pathologic hallmarks of ICI-associated myocarditis observed in patients. Using this model, we show that Ctla4 and Pdcd1 functionally interact in a gene dosage-dependent manner, providing a mechanism by which myocarditis arises with increased frequency in the setting of combination ICI therapy. We demonstrate that intervention with CTLA4-Ig (abatacept) is sufficient to ameliorate disease progression and additionally provide a case series of patients in which abatacept mitigates the fulminant course of ICI myocarditis. SIGNIFICANCE: We provide a preclinical model of ICI-associated myocarditis which recapitulates this clinical syndrome. Using this model, we demonstrate that CTLA4 and PD-1 (ICI targets) functionally interact for myocarditis development and that intervention with CTLA4-Ig (abatacept) attenuates myocarditis, providing mechanistic rationale and preclinical support for therapeutic clinical studies.See related commentary by Young and Bluestone, p. 537.This article is highlighted in the In This Issue feature, p. 521.

Wei SC
Meijers WC
Axelrod ML
Anang NAS
Screever EM
Wescott EC
Johnson DB
Whitley E
Lehmann L
Courand PY
Mancuso JJ
Himmel LE
Lebrun-Vignes B
Wleklinski MJ
Knollmann BC
Srinivasan J
Li Y
Atolagbe OT
Rao X
Zhao Y
Wang J
Ehrlich LIR
Sharma P
Salem JE
Balko JM
Moslehi JJ
Allison JP

Animals
Biomarkers, Tumor
Cardiotoxicity
Disease Management
Disease Models, Animal
Disease Susceptibility
Electrocardiography
Humans
Immune Checkpoint Inhibitors
Mice
Molecular Targeted Therapy
Myocarditis
Neoplasms

Journal Article
Research Support, N.I.H., Extramural


Frontiers in cancer immunotherapy-a symposium report

Abstract: Cancer immunotherapy has dramatically changed the approach to cancer treatment. The aim of targeting the immune system to recognize and destroy cancer cells has afforded many patients the prospect of achieving deep, long-term remission and potential cures. However, many challenges remain for achieving the goal of effective immunotherapy for all cancer patients. Checkpoint inhibitors have been able to achieve long-term responses in a minority of patients, yet improving response rates with combination therapies increases the possibility of toxicity. Chimeric antigen receptor T cells have demonstrated high response rates in hematological cancers, although most patients experience relapse. In addition, some cancers are notoriously immunologically "cold" and typically are not effective targets for immunotherapy. Overcoming these obstacles will require new strategies to improve upon the efficacy of current agents, identify biomarkers to select appropriate therapies, and discover new modalities to expand the accessibility of immunotherapy to additional tumor types and patient populations.

Annals of the New York Academy of Sciences

Cable J
Greenbaum B
Pe'er D
Bollard CM
Bruni S
Griffin ME
Allison JP
Wu CJ
Subudhi SK
Mardis ER
Brentjens R
Sosman JA
Cemerski S
Zavitsanou AM
Proia T
Egeblad M
Nolan G
Goswami S
Spranger S
Mackall CL

Biomarkers, Tumor
Cancer Vaccines
Combined Modality Therapy
Humans
Immune Checkpoint Inhibitors
Immunotherapy
Immunotherapy, Adoptive
Neoplasm Recurrence, Local
Neoplasms
Tumor Microenvironment


Neoadjuvant PD-L1 plus CTLA-4 blockade in patients with cisplatin-ineligible operable high-risk urothelial carcinoma

Abstract: Immune checkpoint therapy is being tested in the neoadjuvant setting for patients with localized urothelial carcinoma1,2, with one study reporting data in cisplatin-ineligible patients who received anti-PD-L1 monotherapy2. The study reported that patients with bulky tumors, a known high-risk feature defined as greater than clinical T2 disease, had fewer responses, with pathological complete response rate of 17%2. Here we report on the first pilot combination neoadjuvant trial ( NCT02812420 ) with anti-PD-L1 (durvalumab) plus anti-CTLA-4 (tremelimumab) in cisplatin-ineligible patients, with all tumors identified as having high-risk features (n = 28). High-risk features were defined by bulky tumors, variant histology, lymphovascular invasion, hydronephrosis and/or high-grade upper tract disease3-5. The primary endpoint was safety and we observed 6 of 28 patients (21%) with grade ≥3 immune-related adverse events, consisting of asymptomatic laboratory abnormalities (n = 4), hepatitis and colitis (n = 2). We also observed pathological complete response of 37.5% and downstaging to pT1 or less in 58% of patients who completed surgery (n = 24). In summary, we provide initial safety, efficacy and biomarker data with neoadjuvant combination anti-PD-L1 plus anti-CTLA-4, which warrants further development for patients with localized urothelial carcinoma, especially cisplatin-ineligible patients with high-risk features who do not currently have an established standard-of-care neoadjuvant treatment.

Gao J
Navai N
Alhalabi O
Siefker-Radtke A
Campbell MT
Tidwell RS
Guo CC
Kamat AM
Matin SF
Araujo JC
Shah AY
Msaouel P
Corn P
Wang J
Papadopoulos JN
Yadav SS
Blando JM
Duan F
Basu S
Liu W
Shen Y
Zhang Y
Macaluso MD
Wang Y
Chen J
Zhang J
Futreal A
Dinney C
Allison JP
Goswami S
Sharma P

Clinical Trial
Journal Article
Research Support, Non-U.S. Gov't


Heterogeneous antibodies against SARS-CoV-2 spike receptor binding domain and nucleocapsid with implications for COVID-19 immunity

Abstract: Evaluation of potential immunity against the novel severe acute respiratory syndrome (SARS) coronavirus that emerged in 2019 (SARS-CoV-2) is essential for health, as well as social and economic recovery. Generation of antibody response to SARS-CoV-2 (seroconversion) may inform on acquired immunity from prior exposure, and antibodies against the SARS-CoV-2 spike protein receptor binding domain (S-RBD) are speculated to neutralize virus infection. Some serology assays rely solely on SARS-CoV-2 nucleocapsid protein (N-protein) as the antibody detection antigen; however, whether such immune responses correlate with S-RBD response and COVID-19 immunity remains unknown. Here, we generated a quantitative serological ELISA using recombinant S-RBD and N-protein for the detection of circulating antibodies in 138 serial serum samples from 30 reverse transcription PCR-confirmed, SARS-CoV-2-hospitalized patients, as well as 464 healthy and non-COVID-19 serum samples that were collected between June 2017 and June 2020. Quantitative detection of IgG antibodies against the 2 different viral proteins showed a moderate correlation. Antibodies against N-protein were detected at a rate of 3.6% in healthy and non-COVID-19 sera collected during the pandemic in 2020, whereas 1.9% of these sera were positive for S-RBD. Approximately 86% of individuals positive for S-RBD-binding antibodies exhibited neutralizing capacity, but only 74% of N-protein-positive individuals exhibited neutralizing capacity. Collectively, our studies show that detection of N-protein-binding antibodies does not always correlate with presence of S-RBD-neutralizing antibodies and caution against the extensive use of N-protein-based serology testing for determination of potential COVID-19 immunity.

McAndrews KM
Dowlatshahi DP
Dai J
Becker LM
Hensel J
Snowden LM
Leveille JM
Brunner MR
Holden KW
Hopkins NS
Harris AM
Kumpati J
Whitt MA
Lee JJ
Ostrosky-Zeichner LL
Papanna R
LeBleu VS
Allison JP
Kalluri R

Adaptive Immunity
Antibodies, Neutralizing
Antibodies, Viral
Betacoronavirus
COVID-19
COVID-19 Testing
Clinical Laboratory Techniques
Coronavirus Infections
Female
Humans
Male
Middle Aged
Nucleocapsid
Outcome Assessment, Health Care
Pandemics
Pneumonia, Viral
Protein Binding
SARS-CoV-2
Sensitivity and Specificity
Seroconversion
Serologic Tests
Spike Glycoprotein, Coronavirus


ARID1A mutation plus CXCL13 expression act as combinatorial biomarkers to predict responses to immune checkpoint therapy in mUCC

Abstract: Immune checkpoint therapy (ICT) can produce durable antitumor responses in metastatic urothelial carcinoma (mUCC); however, the responses are not universal. Despite multiple approvals of ICT in mUCC, we lack predictive biomarkers to guide patient selection. The identification of biomarkers may require interrogation of both the tumor mutational status and the immune microenvironment. Through multi-platform immuno-genomic analyses of baseline tumor tissues, we identified the mutation of AT-rich interactive domain-containing protein 1A (ARID1A) in tumor cells and expression of immune cytokine CXCL13 in the baseline tumor tissues as two predictors of clinical responses in a discovery cohort (n = 31). Further, reverse translational studies revealed that CXCL13-/- tumor-bearing mice were resistant to ICT, whereas ARID1A knockdown enhanced sensitivity to ICT in a murine model of bladder cancer. Next, we tested the clinical relevance of ARID1A mutation and baseline CXCL13 expression in two independent confirmatory cohorts (CheckMate275 and IMvigor210). We found that ARID1A mutation and expression of CXCL13 in the baseline tumor tissues correlated with improved overall survival (OS) in both confirmatory cohorts (CheckMate275, CXCL13 data, n = 217; ARID1A data, n = 139, and IMvigor210, CXCL13 data, n = 348; ARID1A data, n = 275). We then interrogated CXCL13 expression plus ARID1A mutation as a combination biomarker in predicting response to ICT in CheckMate275 and IMvigor210. Combination of the two biomarkers in baseline tumor tissues suggested improved OS compared to either single biomarker. Cumulatively, this study revealed that the combination of CXCL13 plus ARID1A may improve prediction capability for patients receiving ICT.

Goswami S
Chen Y
Anandhan S
Szabo PM
Basu S
Blando JM
Liu W
Zhang J
Natarajan SM
Xiong L
Guan B
Yadav SS
Saci A
Allison JP
Galsky MD
Sharma P

Animals
Biomarkers
Biomarkers, Tumor
Chemokine CXCL13
DNA-Binding Proteins
Humans
Mice
Mutation
Transcription Factors
Tumor Microenvironment
Urinary Bladder Neoplasms


Spatially resolved analyses link genomic and immune diversity and reveal unfavorable neutrophil activation in melanoma

Abstract: Complex tumor microenvironmental (TME) features influence the outcome of cancer immunotherapy (IO). Here we perform immunogenomic analyses on 67 intratumor sub-regions of a PD-1 inhibitor-resistant melanoma tumor and 2 additional metastases arising over 8 years, to characterize TME interactions. We identify spatially distinct evolution of copy number alterations influencing local immune composition. Sub-regions with chromosome 7 gain display a relative lack of leukocyte infiltrate but evidence of neutrophil activation, recapitulated in The Cancer Genome Atlas (TCGA) samples, and associated with lack of response to IO across three clinical cohorts. Whether neutrophil activation represents cause or consequence of local tumor necrosis requires further study. Analyses of T-cell clonotypes reveal the presence of recurrent priming events manifesting in a dominant T-cell clonotype over many years. Our findings highlight the links between marked levels of genomic and immune heterogeneity within the physical space of a tumor, with implications for biomarker evaluation and immunotherapy response.

Mitra A
Andrews MC
Roh W
De Macedo MP
Hudgens CW
Carapeto F
Singh S
Reuben A
Wang F
Mao X
Song X
Wani K
Tippen S
Ng KS
Schalck A
Sakellariou-Thompson DA
Chen E
Reddy SM
Spencer CN
Wiesnoski D
Little LD
Gumbs C
Cooper ZA
Burton EM
Hwu P
Davies MA
Zhang J
Bernatchez C
Navin N
Sharma P
Allison JP
Wargo JA
Yee C
Tetzlaff MT
Hwu WJ
Lazar AJ
Futreal PA

Biomarkers, Tumor
DNA Copy Number Variations
Genomics
Humans
Melanoma
Mutation
Neutrophil Activation
Tumor Microenvironment


Neoantigen responses, immune correlates, and favorable outcomes after ipilimumab treatment of patients with prostate cancer

Abstract: Tumors with high mutational burden (TMB) tend to be responsive to immune checkpoint blockade (ICB) because there are neoantigens available for targeting by reinvigorated T cells, whereas those with low TMB demonstrate limited clinical responses. To determine whether antigen-specific T cell responses can be elicited after treatment with ICB in cancers that have a low TMB, we conducted a clinical trial with ipilimumab in 30 patients with metastatic castration-resistant prostate cancer. We identified two distinct cohorts by survival and progression times: "favorable" (n = 9) and "unfavorable" (n = 10). Patients in the favorable cohort had high intratumoral CD8 T cell density and IFN-γ response gene signature and/or antigen-specific T cell responses. Two patients with a relatively low TMB had T cell responses against unique neoantigens. Moreover, six of nine patients in the favorable group are still alive at the time of analysis, with survival ranging from 33 to 54 months after treatment. All 10 patients in the unfavorable cohort have succumbed to their disease and had survival ranging from 0.6 to 10.3 months. Collectively, our data indicate that immunological correlates associated with effector T cell responses are observed in patients with metastatic prostate cancer who benefit from ICB.

Subudhi SK
Vence L
Zhao H
Blando J
Yadav SS
Xiong Q
Reuben A
Aparicio A
Corn PG
Chapin BF
Pisters LL
Troncoso P
Tidwell RS
Thall P
Wu CJ
Zhang J
Logothetis CL
Futreal A
Allison JP
Sharma P

Antineoplastic Agents, Immunological
Biomarkers, Tumor
CD8-Positive T-Lymphocytes
Cohort Studies
Humans
Ipilimumab
Male
Prostatic Neoplasms


Comprehensive T cell repertoire characterization of non-small cell lung cancer

Abstract: Immunotherapy targeting T cells is increasingly utilized to treat solid tumors including non-small cell lung cancer (NSCLC). This requires a better understanding of the T cells in the lungs of patients with NSCLC. Here, we report T cell repertoire analysis in a cohort of 236 early-stage NSCLC patients. T cell repertoire attributes are associated with clinicopathologic features, mutational and immune landscape. A considerable proportion of the most prevalent T cells in tumors are also prevalent in the uninvolved tumor-adjacent lungs and appear specific to shared background mutations or viral infections. Patients with higher T cell repertoire homology between the tumor and uninvolved tumor-adjacent lung, suggesting a less tumor-focused T cell response, exhibit inferior survival. These findings indicate that a concise understanding of antigens and T cells in NSCLC is needed to improve therapeutic efficacy and reduce toxicity with immunotherapy, particularly adoptive T cell therapy.

Reuben A
Zhang J
Chiou SH
Gittelman RM
Li J
Lee WC
Fujimoto J
Behrens C
Liu X
Wang F
Quek K
Wang C
Kheradmand F
Chen R
Chow CW
Lin H
Bernatchez C
Jalali A
Hu X
Wu CJ
Eterovic AK
Parra ER
Yusko E
Emerson R
Benzeno S
Vignali M
Wu X
Ye Y
Little LD
Gumbs C
Mao X
Song X
Tippen S
Thornton RL
Cascone T
Snyder A
Wargo JA
Herbst R
Swisher S
Kadara H
Moran C
Kalhor N
Zhang J
Scheet P
Vaporciyan AA
Sepesi B
Gibbons DL
Robins H
Hwu P
Heymach JV
Sharma P
Allison JP
Baladandayuthapani V
Lee JJ
Davis MM
Wistuba II
Futreal PA
Zhang J

Adult
Aged
Carcinoma, Non-Small-Cell Lung
Clone Cells
ErbB Receptors
Female
Humans
Lung
Lung Neoplasms
Lymphocyte Activation
Lymphocyte Count
Male
Middle Aged
Mutation
Survival Analysis
T-Lymphocytes


Dissecting the mechanisms of immune checkpoint therapy

Antineoplastic Combined Chemotherapy Protocols


An iPSC-derived neural progenitor cell therapy for subacute spinal cord injury: a phase 1 trial with long-term follow-up

Abstract: Spinal cord injury (SCI) causes irreversible motor and sensory deficits, and no therapy currently restores the damaged neural circuitry. Previous work has shown that transplantation of induced pluripotent stem cell (iPSC)-derived neural stem/progenitor cells (NS/PCs) restored motor function in preclinical models of subacute SCI; however, the safety of this approach in humans remains unknown. Here we report results from a first-in-human, open-label study of iPSC-NS/PC transplantation in four patients with subacute cervical complete SCI. The primary safety end point was achieved, with no tumor formation or graft-related adverse events observed during 2-4 years of follow-up and stable graft sites on imaging. Exploratory efficacy was assessed as a secondary end point. Median improvement in the International Standards for Neurological Classification of Spinal Cord Injury motor score from baseline (2 weeks after injury) to week 52 was 13 points (range 10-40), with two patients improving in American Spinal Injury Association Impairment Scale (grade A → C and A → D). These gains were numerically greater than spontaneous recovery observed in a registry-based cohort. This study provides clinical evidence that transplantation of human iPSC-NS/PCs into the injured spinal cord is feasible and safe under short-term immunosuppression, with findings supporting further clinical evaluation. The trial is registered at UMIN000035074 , UMIN000050104 and jRCTa031190228 .

Sugai K
Tsuji O
Fujiyoshi K
Ozaki M
Abe T
Kohzuki T
Okawara H
Sawada T
Sumida M
Yoshioka E
Shofuda T
Kawashima S
Yamaguchi R
Shinozaki M
Takasu N
Nagoshi N
Okubo T
Kohyama J
Maeda T
Yato Y
Kanemura Y
Yamanaka S
Nakamura M
Okano H


Gonocytes in Transition: Establishing the Male Germline Identity

Abstract: Male germline development is characterized by a unique perinatal phase in which primordial germ cells (PGCs) differentiate into gonocytes (prospermatogonia) and subsequently give rise to spermatogonia. This chapter outlines the cellular, molecular, and epigenetic programs that orchestrate this transition, establishing the foundation of the lifelong spermatogenic lineage. Gonocytes undergo a transient G0/G1 arrest tightly regulated by retinoic acid (RA) metabolism, CDK inhibition, TGF family signaling, and the RNA-binding protein NANOS2, which integrates somatic cues to suppress meiosis and enforce male fate. Concurrently, gonocytes experience genome-wide de novo DNA methylation driven by NSD1-mediated H3K36me2 deposition and the DNMT3A/DNMT3L/DNMT3C machinery, accompanied by piRNA-directed transposon silencing and paternal imprint establishment. These processes collectively reshape the chromatin landscape and stabilize the male germline epigenome. The subsequent gonocyte-to-spermatogonia transition (GST) involves FGF, GDNF, and RA signaling, as well as dynamic histone demethylation, to generate spermatogonial stem cells (SSCs). Aberrant regulation of these pathways can arrest differentiation and predispose germ cells to transformation, as seen in testicular germ cell tumors (TGCTs), whose precursor cells retain gonocyte-like features. We further discuss the interplay between transposon control, imprinting, and chromatin architecture that underpins gonocyte identity. Emerging single-cell and small-input omics approaches are now redefining this transient developmental state, providing new insight into how epigenetic reprogramming and signaling convergence establish the male germline.

Advances in experimental medicine and biology

Li P
Ohhata T
Sakai S
Niida H
Yamanaka S

Gene Expression Regulation, Developmental


Re: Xenotransplantation of a Porcine Kidney for End-stage Kidney Disease

Yamamoto S
Yamanaka S
Yokoo T
Kimura T
Egawa S


Rupture of a Splenic Artery Aneurysm After Thrombolysis Mimicking a Pancreatic Cyst

Circulation journal : official journal of the Japanese Circulation Society

Kobayashi N
Tokuyama H
Yamanaka S
Kanamori T
Sato N


Progressive Giant Left Atrial Appendage Thrombus After Aborted Appendage Occlusion Despite Anticoagulation

Abstract: Thrombus formation after successful left atrial appendage occlusion (LAAO) has been well described; however, the clinical course after aborted LAAO remains poorly characterized. An 84-year-old man with atrial fibrillation and a history of cardioembolic events underwent an attempted LAAO procedure that was aborted because of unfavorable appendage anatomy. A small thrombus suspected on transthoracic echocardiography the day after the procedure progressively enlarged over 3 months and ultimately required surgical thrombus removal and left atrial appendage resection. Thrombus formation after an aborted LAAO attempt is not routinely observed, particularly under therapeutic anticoagulation. In this case, temporary interruption of anticoagulation, local endothelial disruption, blood stasis, and a possible prothrombotic milieu may have contributed. Progressive native left atrial appendage thrombus may occur after aborted LAAO despite anticoagulation therapy. Careful postprocedural imaging surveillance should be considered in high-risk patients.

Kawahara R
Tokuyama H
Yamanaka S
Yui Y
Kobayashi N
Ota H
Miyauchi Y


Effect of a Clinical Decision Support System on Team-Based Clinical Quiz Performance Among Japanese Medical Students: A Pre-Post Pilot Study

Abstract: Educational impact of point-of-care clinical decision support system (CDSS) on medical students remains insufficiently studied. Our aim was to evaluate the effect of a CDSS on team-based clinical quiz performance and confidence in using medical information resources. We conducted a nationwide, online, pre-post quasi-experimental pilot study on November 24, 2024. Eighteen teams of Japanese medical students (team size ranging from one to three members) completed ten multiple-choice questions based on two clinical cases. Teams first answered questions using any resources except UpToDate (Pre phase), then re-answered the same questions using UpToDate only (Post phase) under stricter time constraints. The primary outcome was team-based quiz score. Secondary outcomes were team-based confidence in using UpToDate and other resources. Paired t-tests and Wilcoxon signed-rank tests were performed. Sensitivity analyses addressed missing post-intervention confidence data. Seventeen teams (43 individuals) were included in the analysis. Mean quiz scores increased from 4.00 (SD 1.70) to 4.76 (SD 2.17) (paired t-test p = 0.01; Wilcoxon p = 0.026). Confidence in using UpToDate improved from 1.38 (SD 0.77) to 2.54 (SD 0.97) (paired t-test p = 0.0021), while confidence in other resources showed no significant change. Sensitivity analyses supported the confidence improvement under neutral and best-case assumptions but not under the extreme worst-case scenario. CDSS use may be associated with modest improvements in team-based quiz performance and confidence among medical students in this volunteer-based sample; generalizability to broader populations requires further investigation. Future studies should examine learning outcomes, clinical applicability, and curricular integration.

Isoda S
Hashimoto T
Nishizawa T
Sekiya S
Yamanaka S
Misawa M
Watari T
Ueda E
Suzuki T
Tokuda Y


Transposable element regulation in mammalian germ cells and other contexts: multilayered repression, fertility, and regulated activation

Abstract: Transposable elements (TEs), particularly retrotransposons that dominate mammalian genomes, are pervasive components of mammalian genomes whose activation is constrained by multilayered repression systems. In germ cells, this repression architecture is particularly elaborate, integrating chromatin-based silencing, DNA methylation, and small RNA-guided pathways to safeguard genome integrity during epigenetic reprogramming. These mechanisms are coordinated yet mechanistically specialized, targeting distinct phases of the transposon life cycle and different TE families across developmental stages. Yet the distinctive chromatin landscape of germ cells also creates windows of developmental permissiveness during which TE transcription can occur. Beyond the germline, TE expression can emerge in defined stages of early embryogenesis, extraembryonic development, neural differentiation, and aging, with consequences ranging from chromatin remodeling and regulatory co-option to inflammatory signaling and genome instability. Together, these observations raise a central question: how do different mammalian lineages balance epigenetic plasticity with genome defense? Here, we synthesize current understanding of the molecular logic of TE repression, emphasizing the germline, and integrate evidence across development and aging. We highlight shared principles-such as epigenetic permissiveness and RNA-guided targeting-while underscoring a key difference in regulatory outcome: somatic contexts may tolerate, co-opt, or pathologically amplify TE activity, whereas the germline converts transient activation into heritable, sequence-specific silencing.


Dual TERT promoter mutations in glioblastoma, IDH-wildtype: a case report with preclinical investigations

Abstract: Telomerase reverse transcriptase promoter (TERTp) mutations c.-124C > T (C228T) and c.-146C > T (C250T) are the most frequent single-nucleotide variants promoting telomerase activation in glioblastoma. Here, we describe a rare case of glioblastoma IDH-wildtype with dual TERTp mutations. Genomic analyses of multiregional tumor samples revealed co-occurring TERT C228T and C250T mutations across tumor regions, with variable copy number alterations of EGFR and CDKN2A. Notably, only the TERTp C228T mutation was retained in the corresponding patient-derived xenograft, accompanied by more extensive copy number alterations, implying a selective growth advantage for the TERT C228T-harboring subclone during tumor propagation in this case. These findings suggest that dual TERTp mutations may arise through subclonal evolutionary processes in glioblastoma and highlight telomere maintenance as a dynamic, heterogeneous process rather than invariably representing a fixed early oncogenic event.

Miyake Y
Tateishi K
Okagawa H
Koyama A
Hirata E
Sasaoka K
Saito S
Fushimi S
Natsumeda M
Muraoka E
Yamanaka S
Fujii S
Yamamoto T


Absence of chlamydial infection in ocular adnexal MALT lymphoma based on next-generation sequencing

Abstract: Extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT) arises in various extranodal sites and has been linked to chronic inflammation, often triggered by infectious agents. Ocular adnexal (OA) MALT lymphoma has been associated with Chlamydophila psittaci (C. psittaci) in some regions. This study aimed to investigate the involvement of pathogenic microorganisms, including C. psittaci, in OA-MALT lymphoma. DNA was extracted from 17 formalin-fixed, paraffin-embedded OA-MALT lymphoma samples. Next-generation sequencing was performed, and reads were analyzed with bioinformatic pipelines to detect non-human DNA, focusing on complete or partial genomes of known pathogenic microorganisms, including C. psittaci strain 6BC and other bacteria implicated in lymphomagenesis. Most sequencing reads mapped to the human genome. No complete or partial genomes of C. psittaci 6BC or other non-human pathogenic microorganisms were detected above background levels, and no consistent enrichment of microbial sequences was observed. These findings suggest that, unlike in some Western cohorts, infectious agents such as C. psittaci are unlikely to play a major etiologic role in Japanese OA-MALT lymphoma. Our analysis suggests that infectious agents, including C. psittaci, may be not associated with OA-MALT lymphoma in Japan.

Yakushijin Y
Horiba K
Hashino M
Yamanaka S
Hasebe S
Yamamoto S
Fujii T
Takenaka K
Yasukawa M


eIF4G2-mediated selective translation of chromatin regulators safeguards adult intestinal stem cell identity and differentiation

Abstract: eIF4G2 (DAP5/NAT1) is a non-canonical translation initiation factor, but its role in homeostasis is unclear. Using inducible Eif4g2 knockout mice and intestinal organoids, we show that eIF4G2 loss collapses Lgr5+ intestinal stem cell (ISC) and secretory maturation programs while preserving villus architecture. Transcriptomic and single-nucleus multiome analyses reveal a durable fetal-like/regenerative state with YAP-TEAD activation and regenerative absorptive cells. Ribosome profiling identifies selective translation-efficiency loss among chromatin regulators, especially the KAT3 coactivators CREBBP and EP300, resulting in reduced KAT3 abundance and global histone acetylation; chemical KAT3 inhibition phenocopies this state. CUT&Tag and assay for transposase-accessible chromatin sequencing (ATAC-seq) demonstrate that reduced eIF4G2-KAT3 output drives locus-selective enhancer remodeling, with loss of adult ISC/Wnt-Notch elements and activation of TEAD-enriched fetal loci, without inflammatory or integrated stress response programs driving the transition. Fetal intestinal spheroids remain viable despite similar biochemical defects, highlighting a stage-specific requirement for translational buffering in maintaining adult identity.

Kunitomi H
Khaine AM
Jamee R
Arreola V
Lancero M
Raychaudhuri A
Perli S
Sato Y
Iwasaki M
Ruivo P
Tomoda K
Mito M
Shichino Y
Iwasaki S
Yamanaka S

Animals
Cell Differentiation
Intestines
Mice
Chromatin
Eukaryotic Initiation Factor-4G
Adult Stem Cells
Protein Biosynthesis
Mice, Knockout
Histones
E1A-Associated p300 Protein


Orthognathic surgery and postoperative nausea and vomiting: An 11-year retrospective observational study

Abstract: Orthognathic surgery (OGS) is now widely performed, and postoperative nausea and vomiting (PONV) remains a common complication. Gastric decompression using a nasogastric tube (NT) is generally considered an effective prophylactic measure. In this retrospective observational study, we reviewed 667 OGS cases in 632 patients treated at our department between 2014 and 2024 to evaluate the role of NT placement in PONV by examining associations between clinical variables and PONV. Variables were extracted from electronic medical records, and descriptive statistics and annual trends were summarized. Univariable and multivariable logistic regression analyses were performed to examine associations with PONV. The mean age was 27.2 years, and 462 cases (69.3%) involved female patients. Single-jaw surgery was performed in 363 cases (54.4%) and double-jaw surgery in 304 cases (45.6%). Postoperative NT placement was used in 278 cases (41.7%), and PONV occurred in 53 cases (7.9%). The annual number of OGS cases increased from 48 in 2014 to 123 in 2024. Multivariable logistic regression analysis showed that age ≥ 30 years (odds ratio (OR), 0.43; 95% confidence interval (CI), 0.19-0.95; p = 0.04) and ondansetron administration (OR, 0.21; 95% CI, 0.04-0.97; p = 0.05) were associated with a lower risk of PONV, whereas postoperative NT placement was associated with a higher risk (OR, 4.03; 95% CI, 1.74-9.32; p = 0.001). These findings suggest that the number of OGS cases may be increasing, possibly reflecting growing societal awareness. In addition, postoperative NT placement may increase the risk of PONV, particularly in younger patients, while ondansetron may be effective in reducing PONV.

Watanabe T
Uozumi R
Kawamura T
Sasaki M
Okada R
Inoue S
Kashiwagi M
Fukuhara S
Yamanaka S
Mishima S
Yamaguchi A
Hirota M

Humans
Female
Retrospective Studies
Postoperative Nausea and Vomiting
Adult
Male
Young Adult
Orthognathic Surgical Procedures
Middle Aged
Adolescent
Intubation, Gastrointestinal


Activities of Daily Living Level is Associated With Recovery of Catheter-Free Urination Among Patients Hospitalized in an Acute Care Hospital

Abstract: To examine the Activities of Daily Living (ADL) level required to achieve catheter-free urination in cases where a urinary catheter was inserted due to urinary retention, urination difficulty, or urinary incontinence in an acute care hospital. This study included patients who were originally capable of self-urination but required rehabilitative therapy and intervention by the multidisciplinary urination care team for difficulty in removal of indwelled Foley catheter during hospitalization. The primary outcome was catheter-free independent urination. Patients who achieved urinary independence through self-catheterization for prolonged neurogenic bladder dysfunction were excluded. ADL scores before and after the intervention were evaluated using the Functional Independence Measure (FIM), which was consisted of motor FIM (mFIM), cognitive FIM, and total FIM, with and without deduction for urinary control score (-UC). Logistic regression analyses were performed to identify factors associated with independent urination, and Receiver Operating Characteristic (ROC) curve analysis was conducted to determine the appropriate cutoff. The study included 87 cases (72.8 ± 12.4 years old, male:female = 51:36). Catheter-free urination was achieved in 43 (49%) patients. There were no significant differences between the catheter-dependent and catheter-free groups in terms of gender, age, hospitalized departments, treatment methods, total duration of admission, duration of intervention, discharged destination, and comorbidities. At the end of the intervention, the FIM scores were all significantly higher in the catheter-free group than in the catheter-dependent group. To achieve catheter-free urination, mFIM-UC at the end of the intervention was the only statistically significant corelate (p < 0.001). In the ROC analysis, the AUC of mFIM-UC at the end of the intervention was 0.834, and the optimal cutoff value was 30 points (full score 84), with a sensitivity of 0.91 and specificity of 0.66. ADL as represented by mFIM is a major correlate of catheter-free urination.

Tanaka T
Kanematsu A
Nagai N
Yoneda H
Yamanaka S
Kou Y
Yanagi T
Yamada Y
Saito R
Domen K
Yamamoto S

Humans
Female
Male
Activities of Daily Living
Urinary Catheterization
Recovery of Function
Aged
Aged, 80 and over
Middle Aged
Urination
Urinary Retention
Hospitalization
ROC Curve
Treatment Outcome
Urinary Catheters
Urinary Incontinence
Logistic Models


Effects of different mycorrhizal types of dispersedly retained trees on the diversity of ectomycorrhizal fungi in neighboring Abies sachalinensis seedlings

Abstract: Retention forestry conserves biodiversity by retaining forest structures in logged areas. It has been demonstrated that dispersedly retained broad-leaved ectomycorrhizal (EcM) trees can mitigate the effect of logging on the diversity of EcM fungi in the surrounding Abies sachalinensis seedlings. However, it remains unclear how retained trees of different mycorrhizal types affect the diversity of EcM fungi in Abies seedlings. We investigated the neighborhood effect of different mycorrhizal types of retained trees on the diversity of EcM fungi symbiotic with surrounding Abies seedlings. At dispersed retention sites, the roots of Abies seedlings were collected near mature EcM trees (ET) or arbuscular mycorrhizal (AM) trees (AT), or in open areas where no retained trees existed within ten meters (NT). EcM fungi were identified based on ITS barcoding of the EcM roots. The diversity measures of EcM fungi under AT and NT were comparable and lower, respectively than those under ET. The community composition of the EcM fungi was similar between AT and NT, and both were significantly different from that of ET. These results indicate that AM trees do not have significant impact on EcM fungul community in the surrounding EcM seedlings.


Temporal AI model predicts drivers of cell state trajectories across human aging

Abstract: Foundational AI models have recently shown promise for predicting the impact of perturbations on cell states. However, current models typically consider only one cell state at a time, limiting their ability to learn how cellular responses unfold over time, particularly across long trajectories such as diseases of aging. Here, we develop a temporal AI model, MaxToki, trained on nearly 1 trillion gene tokens including cell state trajectories across the human lifespan to generate cell states across long timelapses of human aging. MaxToki generalized to unseen trajectories through in-context learning and predicted novel age-modulating targets that were experimentally verified to influence age-related gene programs and functional decline in vivo. MaxToki represents a promising strategy for temporal modeling to accelerate the discovery of interventions for programming therapeutic cellular trajectories.

bioRxiv : the preprint server for biology

Ortega JG
Nadadur RD
Kunitomi A
Kothen-Hill S
Wagner JUG
Kurtoglu SD
Kim B
Reid MM
Lu T
Washizu K
Zanders L
Chen H
Zhang Y
Ancheta S
Lichtarge S
Johnson WA
Thompson C
Phan DM
Combes AJ
Yang AC
Tadimeti N
Dimmeler S
Yamanaka S
Alexanian M
Theodoris CV


Combined Large Cell Neuroendocrine Carcinoma of the Endometrium Treated With Paclitaxel, Carboplatin, and Durvalumab, Followed by Maintenance Therapy With Durvalumab and Olaparib (DUO-E Regimen): A Case Report

Abstract: Combined large cell neuroendocrine carcinoma (LCNEC) of the endometrium is rare and carries a poor prognosis. A 61-year-old woman with suspected endometrial serous carcinoma was referred to our hospital. Imaging studies staged the disease as stage IIIC1. We performed a total hysterectomy, bilateral salpingo-oophorectomy, omentectomy, and retroperitoneal lymph node dissection. Histopathological examination revealed a combination of LCNEC, endometrioid carcinoma, and multiple lymph node metastases. Postoperative computed tomography showed multiple pulmonary metastases not detected preoperatively. Given the advanced and recurrent nature of the disease, treatment with paclitaxel, carboplatin, and durvalumab was initiated. After the first cycle, the pulmonary metastases disappeared. Maintenance therapy with durvalumab and olaparib was continued because of proficient mismatch repair status. This case demonstrates that durvalumab may be a viable treatment option for endometrial LCNEC, as it is also approved for small cell lung cancer, which shares similarities with neuroendocrine carcinoma.

The journal of obstetrics and gynaecology research

Suzuki T
Sonehara K
Nabeshima H
Shioya Y
Yamanaka S
Kondo S


Factors Associated and Survival Outcomes Among Japanese Patients with Solid Cancer Who Developed Deep Vein Thrombosis (DVT)

Abstract: Deep vein thrombosis (DVT) is a common and serious complication in patients with cancer. We retrospectively analyzed patients newly diagnosed with solid cancers in a Japanese cohort. We retrospectively reviewed the medical records (2013-2020) of Japanese patients with solid cancers who were suspected of having DVT and were diagnosed and treated at Ehime University Hospital. Controls were patients with solid cancers in whom DVT was suspected on physical examination and/or laboratory testing but subsequently ruled out by definitive imaging. Candidate risk and prognostic variables were extracted from electronic medical records. This was a single-center cohort study. Among the initial 1399 patients, 224 with DVT and 560 without DVT (as controls) were included in the final analysis after excluding those with incomplete medical records, missing diagnostic tests, or unclear medical histories. The median overall survival (OS) period was 5.87 years for patients with DVT and 6.81 years for patients without DVT (p=0.0014). Anemia (Hb <11 g/dl) and thrombocytopenia (Plt <15 x 104 / μL) were found to be strong risk factors related to an increased incidence of DVT. Female cancer patients with DVT had a significantly worse outcome than those without DVT (p=0.028). Analysis of OS in patients with DVT associated with gynecological cancers following treatment with three different direct oral anticoagulants (DOACs; edoxaban, rivaroxaban, and apixaban) indicated that those treated with apixaban had a significantly worse outcome than those treated with the others (p=0.019) in this Japanese cohort. Japanese patients with solid cancers, particularly those with gynecologic cancers, tend to have poorer outcomes when DVT co-occurs with anemia and low platelet counts. In addition, the choice of DOAC for DVT treatment may be associated with differences in prognosis. These exploratory findings require more detailed, adjusted analyses and confirmation using other datasets or populations.

International journal of general medicine

Masuda Y
Yakushijin Y
Yamanaka S
Hasebe S
Fujii T
Yamanouchi J
Takenaka K


An interactome-based framework for DDB1- and CUL4-associated factor prioritization in targeted protein degradation

Abstract: The DDB1- and CUL4-associated factor (DCAF) family functions as substrate receptors within Cullin4-really interesting new gene (RING) ubiquitin ligases (CRL4s), facilitating proteasomal degradation of targeted substrates. Although CRL4-based targeted protein degradation (TPD) has emerged as a promising strategy to modulate undruggable proteins, the complex formation, substrates, and functional properties of many DCAFs remain poorly defined. In this study, using proximity biotinylation-based interactome analysis in human HEK293T cells, we systematically annotated interactors and functional associations of individual DCAFs. Furthermore, we identified substrates of the model DCAFs COP1 and DCAF3 using proximity biotinylation coupled with multi-omics approaches. By combining biochemical and cell-based analyses, we establish CRL4 complex formation and DCAF autodegradation as experimentally tractable proxy indicators to evaluate DCAF degradation activity and propose a set of high-activity DCAFs. These datasets establish a resource for functional characterization of DCAFs and provide a framework for their prioritization in TPD.

Yamanaka S
Nagaoka K
Shoya Y
Nishino K
Mikura Y
Tanaka K
Konishi K
Hasegawa Y
Hijikata A
Kosako H
Sawasaki T

Humans
HEK293 Cells
Proteolysis
DNA-Binding Proteins
Cullin Proteins
Ubiquitin-Protein Ligases
Carrier Proteins
Proteasome Endopeptidase Complex
Ubiquitination
Protein Binding
Biotinylation
Protein Interaction Mapping
Protein Interaction Maps
Peptide Termination Factors


Shinya Yamanaka

Abstract: Dr. Shinya Yamanaka is recognized for the generation of induced pluripotent stem cells (iPSCs) from fibroblasts by a combination of multiple transcription factors, and he won the Nobel Prize in Physiology or Medicine in 2012 jointly with Sir John B. Gurdon for this discovery. Twenty years after the discovery, the Cell Reports Medicine editorial team discusses with Dr. Yamanaka the scientific, technical, and translational milestones that have shaped the field of regenerative medicine. We also discuss the role of iPSCs in disease modeling and drug discovery, the interplay with genome editing, and ongoing issues that still prevent the widespread clinical application of iPSC-derived therapies. Finally, Dr. Yamanaka reflects on promising, yet underexplored, applications of iPSCs.

Induced Pluripotent Stem Cells
Humans
History, 21st Century
History, 20th Century
Regenerative Medicine
Animals
Nobel Prize


Alkyne Two-Phase Strategy: Rapid Generation of TK-285-Derived PROTACs as BRD4 Degraders

Abstract: This study introduces a divergent synthetic strategy in linkerology using preassembled linkers to generate structural diversity. The approach was validated by developing bromodomain-containing protein 4 (BRD4)-targeting proteolysis-targeting chimeras (PROTACs) based on an "alkyne two-phase strategy," employing the BRD4 inhibitor TK-285 as the binding ligand. In the initial screening phase, alkyne-modified TK-285 derivatives were subjected to click chemistry to optimize linker length and the modification site, leading to the identification of TKP-5 as a potent degrader. TKP-5 exhibited stronger thymic stromal lymphopoietin─more suppressive activity than TK-285 and markedly suppressed IL-33 mRNA expression in a tape-stripping-induced skin injury model. In the subsequent optimization phase, late-stage diversification using 1,3-butadiyne-typed PROTAC intermediates revealed the critical contribution of the triazole moiety, supported by in silico analysis suggesting interaction with Trp81 of BRD4. The strategy is expected to be broadly applicable to modular functional molecules accessible via click chemistry.

Yamakoshi H
Watanabe R
Segawa R
Ishihara R
Tachibana R
Kudo G
Nagasawa S
Yamanaka S
Ito A
Takeda H
Sawasaki T
Yoshino R
Hirokawa T
Doi T
Hirasawa N
Iwabuchi Y

Bromodomain Containing Proteins
Proteolysis Targeting Chimera
Cell Cycle Proteins
Humans
Transcription Factors
Animals
Alkynes
Click Chemistry
Structure-Activity Relationship
Mice
Triazoles
Proteolysis


BRAF-mutant high-grade glioma with pleomorphic and pseudopapillary features (HPAP): A PLNTY mimic demonstrating tumor progression during longitudinal follow-up

Abstract: High-grade glioma with pleomorphic and pseudopapillary features (HPAP) is a recently recognized glioma subtype defined by DNA methylation profiling. While it exhibits overlapping histological features with various CNS tumors, such as polymorphous low-grade neuroepithelial tumor of the young (PLNTY) and pleomorphic xanthoastrocytoma, its molecular pathogenesis and clinical behavior remain incompletely understood. We report a rare case of HPAP with BRAF p.V600E mutation and PLNTY-like histological features that showed rapid tumor progression during long-term follow-up. A 47-year-old woman harbored a lesion that remained asymptomatic and slow-growing for over 20 years, but later exhibited contrast enhancement and rapid expansion. Partial tumor resection was performed with hippocampal preservation based on intraoperative genetic testing and functional considerations. No regrowth of the residual hippocampal lesion was observed at 12 months postoperatively. Histologically, the tumor showed oligodendroglioma-like morphology, strong CD34 immunopositivity, consistent with PLNTY-like features, but indicated a high proliferative index. Molecular analysis revealed co-occurring BRAF p.V600E and TERT promoter (pTERT, c.-124C>T) mutations, with a lower variant allele frequency for the pTERT mutation. This disparity, confirmed by droplet digital PCR, suggests that the BRAF p.V600E mutation was an early, clonal event, whereas the pTERT mutation likely arose later in a subclonal population. DNA methylation profiling classified the tumor as HPAP with high confidence (NCI-Bethesda score: 0.969), and uniform manifold approximation and projection showed clustering within HPAP reference cases. This case represents a rare example of BRAF p.V600E-mutant HPAP with PLNTY-like features in which a subclonal pTERT mutation likely emerged during tumor evolution, contributing to rapid tumor progression. The combination of a prolonged indolent phase followed by rapid growth, along with the intratumoral genetic heterogeneity observed, provides novel insights into the biological diversity and evolutionary dynamics of HPAP.

Takayama Y
Yaegashi M
Satomi K
Ishiyama T
Shioda M
Yazawa O
Ye W
Okagawa H
Saito S
Sasaoka K
Okura M
Koyama A
Oshima A
Sonoda M
Natsumeda M
Ichimura K
Yamanaka S
Fujii S
Yamamoto T
Tateishi K


Two decades of induced pluripotent stem cell research: From discovery to diverse applications

Abstract: Twenty years have passed since the first demonstration of mouse induced pluripotent stem cells (iPSCs). What began as an unexpected observation in Kyoto quickly transformed stem cell biology and regenerative medicine worldwide. Over the past two decades, we have gained profound insights into the molecular mechanisms underlying cellular reprogramming and pluripotency. The technology has continued to evolve-becoming safer, more efficient, and more versatile. Today, iPSCs serve as a foundation for wide-ranging applications, from disease modeling and drug discovery to regenerative therapies and rejuvenation research. In this review, I reflect on the scientific journey of iPSCs, highlight key milestones in our understanding of reprogramming, and discuss the expanding clinical and societal impact of iPSCs.

Induced Pluripotent Stem Cells
Animals
Humans
Cellular Reprogramming
Regenerative Medicine
Stem Cell Research


Linking the heart-eye-brain axis: Ocular and cerebral blood flow changes after catheter ablation in atrial fibrillation

Yamamoto N
Suzuki H
Nakano M
Yamanaka S
Ito T
Sato H
Chiba T
Nochioka K
Takanami K
Yasuda M
Kunikata H
Nakazawa T
Yasuda S


B-Cell Lymphoma Following an Indolent Course Over 30 Years with Immunophenotypic Changes at Each Recurrence

Abstract: Diffuse large B-cell lymphoma (DLBCL) comprises biologically distinct subtypes, but whether cell-of-origin-related features remain stable over long disease courses is unclear. A woman was initially diagnosed with CD20-positive B-cell lymphoma in 1988 and treated with cyclophosphamide, vincristine and prednisolone (COP) chemotherapy. She experienced a first relapse in 1998 and received cyclophosphamide, doxorubicin, vincristine and prednisolone (CHOP) chemotherapy. The second relapse occurred in 2008 and was managed with rituximab. At the third relapse in 2011, rituximab plus bendamustine (RB) was administered. During the fourth relapse in 2015, R-pirarubicin (THP)-COP chemotherapy was given. At the fifth relapse in 2017, at 80 years of age, RB therapy was administered again. A sixth relapse occurred in 2019, and the patient died within the same year. Histopathological evaluation throughout the disease course consistently demonstrated DLBCL. Notably, the immunophenotype showed sequential changes from a germinal centre B-cell-like pattern to an activated B-cell-like pattern and ultimately to a CD30-positive anaplastic B-cell variant. This unusually long clinical course highlights stepwise immunophenotypic evolution during repeated relapses, suggesting that surrogate subtype profiles may change over time. Repeat biopsies with comprehensive pathological (and, when available, molecular) re-evaluation are important to accurately characterize relapsed DLBCL and inform treatment selection. Diffuse large B-cell lymphoma (DLBCL) can undergo stepwise immunophenotypic and molecular evolution over decades, progressing from a germinal Center B-cell-like (GCB)-like phenotype to an activated B-cell -like/non-GCB pattern (MUM1+, CD30+) and ultimately to an aggressive anaplastic variant.The cell-of-origin profile and its associated molecular features are not necessarily static; thus, therapeutic strategies for relapsed DLBCL should be tailored to the current disease state rather than relying on the signature at initial diagnosis.Longitudinal re-evaluation through repeat biopsies is imperative to capture the evolving landscape of the malignancy, ensuring that treatment selection is guided by the most recent pathological and molecular findings.

European journal of case reports in internal medicine

Yamanaka S
Hasebe S
Fujii T
Gondo T
Kitazawa R
Kitazawa S
Yakushijin Y


Intraoperative Hypotension and Vasoactive Treatment: An International Survey of Anaesthesiologists

Abstract: Intraoperative hypotension is a common occurrence in patients undergoing anaesthesia, although there is no standardised definition of hypotension. International consensus statements provide some guidelines for the management of intraoperative hypotension, but general clinical practice is unknown. We aimed to survey anaesthesiologists' values and preferences regarding intraoperative blood pressure management, including whether they would support future research on this topic. We conducted an international, online survey of routine practice and opinion. The target population was anaesthesiologists who regularly anaesthetise adult patients. Results are reported descriptively and in accordance with the Consensus-Based Checklist for Reporting of Survey Studies (CROSS) checklist. A total of 1640 anaesthesiologists from 11 European countries participated in the survey. The majority of respondents were specialists (1322 of 1640, 80.6%, 95% CI 78.7-82.6). Almost all respondents worked in public hospitals (1613 of 1640, 98.4%). The overall response rate was 22.7%. Most respondents reported using absolute mean arterial pressure as their main unit of measurement to quantify hypotension (1098 of 1640, 67.0%, 95% CI 64.6-69.2). Respondents were most likely to initiate vasoactive treatment at a mean arterial pressure below 60 or 65 mmHg. Chronic arterial hypertension, traumatic brain injury and surgical procedures involving head-up positioning of the patient were the three most common scenarios where respondents would raise their threshold for treatment. Most respondents considered the establishment of safe intraoperative blood pressure thresholds a critical research question, and almost all respondents (1509 of 1640, 92.0%) indicated a willingness to randomise patients to specific blood pressure targets. For 72.9% (1196 of 1640), the lowest acceptable mean arterial pressure for randomisation was 60 mmHg. Respondents were also interested in the comparison of efficacy and safety of vasoactive agents, and the most sought-after comparison was phenylephrine versus noradrenaline (1252 of 1640, 76.3%). The willingness of respondents to administer these agents in peripheral venous access differed according to geography. In this international survey, mean arterial pressures of 60 or 65 mmHg were the most commonly reported blood pressure thresholds leading to initiation of treatment with vasoactive agents. Almost all respondents indicated patient groups for whom they would alter their treatment threshold, namely those suffering from chronic arterial hypertension, those undergoing surgery in a head-up position, and patients with traumatic brain injury. The majority of respondents supported future trials establishing optimal mean arterial pressure threshold and choice of vasoactive agent. We noticed a geographical variation in willingness to administer vasoactive agents in peripheral venous access. This survey of anaesthesiologists from European countries queried practitioner perceptions of blood pressure management in adults during anaesthesia with focus on hypotension. Queries and responses also concerned circumstances and blood pressure levels which clinicians report being willing to treat actively, and how they might do this practically.

Bækgaard ES
Vester-Andersen M
Crone V
Møller MH
Yamanaka S
Palmarsdottir R
Uusalo P
Haidl F
Rådestad M
van der Sloot K
Corona A
Johnström A
Nørskov AK
Karlsen APH
Faustad BI
Sørensen CB
Spies F
Krogh HB
El-Hallak H
Mistry JM
Mønnich JK
Olesen KG
Madsen KPD
Voogd KL
Reich LM
Kolstrup LA
Juhl-Olsen P
Körner LK
Sigurdsson MI
Jensen-Holm MB
Slavensky JA
Toft MH
Jørgensen ML
Andersen M
Jensen MSH
Saei M
Hedetoft M
Hansen PM
Hansen RT
Lundsgaard RS
Krarup SMI
Wiberg S
Laustrup T
Brizzi G
Kreutziger J
Onyemuchara I
Adeniji A
Colville T
Keitley JA
Khan M
Millar M
Lennie I
Kelly K
Brooker V
Roberts J
Lipton G
Keohone J
Chebbout R
Bond O
Milligan W
O'Brien C
Wright E
Linton F
Towell C
Shuttleworth J
Norton J
Butler D
Frankland S
Screech F
Charig L
Walsh S
Davies R
Jones E
Dalmonte E
Evans M
Krag M

Humans
Intraoperative Complications
Hypotension
Vasoconstrictor Agents
Anesthesiologists
Adult
Female
Surveys and Questionnaires
Male
Europe
Middle Aged


Protocol for In Utero Fetal-to-Fetal Kidney Transplantation in Rats

Abstract: Congenital renal disorders, such as the Potter sequence, result from renal dysgenesis. To explore a prenatal therapeutic approach for fetuses with kidney insufficiency, we established an in utero transplantation protocol using donor fetal kidneys. Although numerous rodent studies have reported cellular injections into fetal recipients, no protocol to date has described whole-organ transplantation during gestation. Here, we present a step-by-step method for grafting donor fetal kidneys (embryonic day 14.0-16.5) into allogeneic rat fetuses at embryonic day 18.0-18.5, resulting in term neonates that retain the grafts postnatally. A 15-16 G needle preloaded with the donor kidney is inserted transuterinely, depositing the organ into the subcutaneous space of the fetus. Four days later, the term pups are delivered naturally and evaluated for graft development. This protocol enables organ-level transplantation and longitudinal assessment of graft maturation within the unique fetal environment, which differs markedly from adult settings in terms of growth factor availability and immune reactivity. To our knowledge, this is the first protocol to successfully achieve whole-organ transplantation directly into fetuses in utero. Therefore, the model provides a valuable platform for studying developmental organogenesis, fetal immunology, and regenerative strategies that leverage embryonic cues. Key features • Subcutaneous transplantation of fetal kidneys into recipient fetuses minimizes surgical invasiveness and significantly improves fetal survival. • Natural delivery enables pups to nurse from the dam, allowing extended postnatal observation. • Use of green fluorescent protein (GFP)-expressing donor tissue permits real-time visualization of graft location and growth. • The protocol is readily adaptable for xenotransplantation and studies of immunological tolerance during fetal development.