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PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The uncropped Northern blot image can be found in Supplementary Figure 4. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The distribution of LINC00870 in GIST cell lines. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The sequence protein-coding ability of LINC00870 was predicted using the prediction website. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Agarose gel electrophoresis was utilized to determine the length of LINC00870 in GIST-882 cells overexpressing either the sense or antisense strands.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The uncropped gel image can be found in Supplementary Figure 5. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Western blot analysis was performed to evaluate the protein-coding potential of LINC00870 in GIST-882 cells overexpressing either the sense or antisense strands, with a luciferase plasmid used as the control.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The uncropped Western blot image can be found in Supplementary Figure 6. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The identification of LINC00870 interference and overexpression efficiency. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
H–I) The effect of interfering (H) or overexpressing (I) LINC00870 on GIST cells' invasive and migratory capacity. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
J–K) The effect of interfering (J) or overexpressing (K) LINC00870 on the ability of GIST clone formation.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Mean ± SEM of 3 independent experiments.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
∗∗∗P < 0.001 versus control.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Fig. 5 Basic characteristics and biological functions of LINC00870. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
LINC00870 sequence information and length were analyzed using 3′RACE and 5′RACE experiments.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The uncropped gel image can be found in Supplementary Figure 3. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The number and length of LINC00870 transcripts in GIST cell lines and tissues were determined by a Northern blot assay.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The uncropped Northern blot image can be found in Supplementary Figure 4. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The distribution of LINC00870 in GIST cell lines. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The sequence protein-coding ability of LINC00870 was predicted using the prediction website. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Agarose gel electrophoresis was utilized to determine the length of LINC00870 in GIST-882 cells overexpressing either the sense or antisense strands.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The uncropped gel image can be found in Supplementary Figure 5. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Western blot analysis was performed to evaluate the protein-coding potential of LINC00870 in GIST-882 cells overexpressing either the sense or antisense strands, with a luciferase plasmid used as the control.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The uncropped Western blot image can be found in Supplementary Figure 6. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The identification of LINC00870 interference and overexpression efficiency. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
H–I) The effect of interfering (H) or overexpressing (I) LINC00870 on GIST cells' invasive and migratory capacity. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
J–K) The effect of interfering (J) or overexpressing (K) LINC00870 on the ability of GIST clone formation.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Mean ± SEM of 3 independent experiments.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
∗∗∗P < 0.001 versus control.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Since lncRNA has been shown to play a crucial role in tumorigenesis and progression [, , , ], we constructed LINC00870 knockdown and overexpression stable cell lines in GIST-T1 and GIST-882 cells to explore its function (Fig. 5G).
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
We observed a reduction in cell invasion, migration, and clone formation after LINC00870 knockdown, and an enhancement of these cellular processes after LINC00870 overexpression (Fig. 5H–K).
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
These results suggest that LINC00870 functions as an oncogene in GIST.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
We selected two independent cohorts to explore the correlation between PIGR and LINC00870 in GIST imatinib-resistant patient samples to validate our findings.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
We found a significant positive correlation between PIGR and LINC00870 in two cohorts (Fig. 6A, P < 0.0001).
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Previous studies have shown that PIGR is a transmembrane glycoprotein whose N-terminal glycosylation promotes hydrolysis and secretion in the extracellular compartment, a critical step in the cellular response to the immune response .
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
We investigated the effect of LINC00870 on PIGR by overexpressing it in GIST cells and found a significant decrease in the secretory component content released extracellularly by PIGR (Fig. 6B).
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
This result was consistent with the effect of tunicamycin treatment, which disrupts the extracellular glycan chain of PIGR (Fig. 6C).
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Our findings suggest that LINC00870 may cause imatinib treatment resistance in GIST by inhibiting the N-terminal glycosylation of PIGR, preventing its extracellular SC region from being released typically and thereby hindering the immune response.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Fig. 6Correlation and mechanism study of PIGR and LINC0087.(A) The correlation analysis between PIGR and LINC00870 at the mRNA level in GIST imatinib-resistant patients.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The unadjusted original results can be found in Supplementary Figure 7. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The expression content of PIGR extracellular secretory structure SC in cell supernatants after overexpression of LINC00870. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The expression content of PIGR extracellular secretory structure SC in cell supernatants after treatment of cells with tunicamycin.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Mean ± SEM of 3 independent experiments.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
∗∗∗P < 0.001 versus control.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Fig. 6 Correlation and mechanism study of PIGR and LINC0087. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The correlation analysis between PIGR and LINC00870 at the mRNA level in GIST imatinib-resistant patients.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The unadjusted original results can be found in Supplementary Figure 7. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The expression content of PIGR extracellular secretory structure SC in cell supernatants after overexpression of LINC00870. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The expression content of PIGR extracellular secretory structure SC in cell supernatants after treatment of cells with tunicamycin.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Mean ± SEM of 3 independent experiments.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
∗∗∗P < 0.001 versus control.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
In our final analysis, we demonstrated that inhibition of PIGR or LINC0087 effectively overcomes imatinib resistance.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Subcutaneous tumor models were established in BALB/c-nu nude mice using GIST-882 or GIST-882_PIGR_KO resistant cells.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Our results indicate that knockout of PIGR in resistant cell lines effectively inhibits tumor growth (Fig. 7A and B).
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Similarly, knockout of LINC0087 in resistant cells also significantly suppresses tumor growth (Fig. 7C and D).
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
These findings collectively suggest that inhibition of PIGR or LINC0087 represents a promising strategy for overcoming imatinib resistance.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Fig. 7Inhibition of PIGR or LINC0087 effectively overcomes imatinib resistance.(A–B) Subcutaneous tumor models were established in BALB/c-nu nude mice using GIST-882 or GIST-882_PIGR_KO resistant cells, and tumor growth was monitored (A).
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Tumor weight was measured at the end of the experiment (B). (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
C–D) Subcutaneous tumor models were established in BALB/c-nu nude mice using GIST-882 or GIST-882_LINC0087_KO resistant cells, and tumor growth was monitored (C).
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Tumor weight was measured at the end of the experiment (D).
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Mean ± SEM of 3 independent experiments.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
∗∗∗P < 0.001 versus control.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Fig. 7 Inhibition of PIGR or LINC0087 effectively overcomes imatinib resistance. (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
A–B) Subcutaneous tumor models were established in BALB/c-nu nude mice using GIST-882 or GIST-882_PIGR_KO resistant cells, and tumor growth was monitored (A).
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Tumor weight was measured at the end of the experiment (B). (
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
C–D) Subcutaneous tumor models were established in BALB/c-nu nude mice using GIST-882 or GIST-882_LINC0087_KO resistant cells, and tumor growth was monitored (C).
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Tumor weight was measured at the end of the experiment (D).
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Mean ± SEM of 3 independent experiments.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
∗∗∗P < 0.001 versus control.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
GIST is the most common type of gastrointestinal mesenchymal tumors.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Their pathogenesis is primarily driven by gain-of-function mutations in the KIT gene (accounting for 75–80 %) and mutations in the platelet-derived growth factor receptor alpha (PDGFRA) (<10 %).
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Additionally, abnormalities in genes such as SDH, NF1, KRAS, and HRAS have been implicated in GIST development.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
GIST cells originate from interstitial cells of Cajal or their precursor cells and their survival is dependent on the expression of KIT .
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
KIT mutations predominantly occur in the juxtamembrane (JM) domain (encoded by exon 11) and in the extracellular domain near the membrane (encoded by exons 8 or 9), with approximately 85–90 % of KIT-mutant GIST exhibiting these characteristics.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
These mutations confer oncogenic properties to the KIT gene, promoting tumor formation primarily through the PI3K-AKT, JAK-STAT, and RAS-RAF-MEK-ERK (MAPK) signaling pathways [, , , , ].
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Currently, imatinib remains the first-line targeted therapy for progressive or unresectable GIST.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Nevertheless, both primary and secondary drug resistance frequently arise, leading to diminished therapeutic efficacy, increased tumor recurrence, metastasis, and elevated mortality rates.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Thus, elucidating the molecular mechanisms underlying drug resistance in GIST is essential for enhancing therapeutic strategies and prognostic outcomes.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Approximately 90 % of GIST cases are driven by activating mutations in receptor tyrosine kinases, primarily KIT or PDGFRA .
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Research on imatinib resistance has highlighted several key factors: KIT mutations result in sustained protein activation, which impacts the genomic context and activates alternative signaling pathways .
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Patients with the PDGFRA-D842V mutation exhibit primary resistance to imatinib .
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
For BRAF/KRAS mutations, while imatinib inhibits the mutated KIT receptor, it does not affect the downstream RAS-RAF signaling pathways associated with BRAF/KRAS mutations .
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Additionally, inhibiting KIT signaling can lead to the release of FGF-2, further worsening tumor malignancy .
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
In terms of metabolism, succinate dehydrogenase (SDH) deficiency results in the stabilization and accumulation of HIF1-α under normoxic conditions, promoting angiogenesis and glycolysis, thereby facilitating tumor growth .
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Given the extremely limited treatment options for imatinib resistance, there is an urgent need to investigate new mechanisms to combat this challenge.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Our high-throughput sequencing of tissue specimens from imatinib-resistant GIST patients identified PIGR as a significantly differentially expressed gene.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
While PIGR has been found to exhibit differential expression in various cancers, there is no conclusive evidence regarding its role in the prognosis of tumor patients [, , , , , , , ].
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Our study discovered that PIGR is remarkably highly expressed in GIST tumor tissues.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
The role of PIGR in tumors remains controversial, possibly owing to tumor heterogeneity or different action mechanisms.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Studies in colorectal cancer revealed that PIGR could act as a prognostic marker and inhibit tumorigenesis through LAMB3-AKT-FOXO3/4 signaling .
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Transcriptome sequencing analysis revealed that PIGR was downregulated in breast cancer, and overexpression of PIGR in breast cancer cells inhibited their adhesion and proliferation .
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
PIGR controlled tumor progression in lung cancer by downregulating the differentiation suppressor gene NOTCH3 .
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
In hepatocellular carcinoma, extracellular vesicles carrying PIGR activated the AKT/GSK3β/β-linked protein signaling cascade response, promoting cancer stemness, tumorigenesis, and metastasis .
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Other studies have revealed that PIGR promoted hepatocellular carcinoma by activating Smad and Yes-MEK/ERK signaling pathways .
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Ohkuma et al. found that PIGR overexpression was associated with poor prognosis after surgical resection in patients with pancreatic cancer .
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Our study found that GIST patients with high PIGR expression had a poor prognosis, and overexpression of PIGR significantly promoted GIST cell invasion, lung colonization, and liver metastasis, suggesting that PIGR functions as an oncogene in GIST.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Our animal experiments also confirmed that interfering with PIGR expression in imatinib-resistant cell lines helps inhibit tumor growth, suggesting that the application of PIGR inhibitors in clinical settings may improve survival in imatinib-resistant patients.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
However, PIGR plays a critical role in mediating the transcytosis of polymeric IgA and IgM from the basolateral surface to the apical surface of epithelial cells, subsequently secreting these immunoglobulins into the mucosal fluid [, , ].
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Therefore, the sole use of PIGR inhibitors could potentially disrupt normal mucosal immunity.
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
If it is possible to specifically interfere with PIGR at tumor sites, for example, through the use of antibody-drug conjugates (ADCs), it may achieve better therapeutic outcomes while reducing side effects for patients .
PMC11834037
LINC00870 promotes imatinib resistance in gastrointestinal stromal tumor via inhibiting PIGR glycosylation modifications
Of course, these conclusions are based solely on animal models, and further challenges must be addressed before clinical application.