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To investigate the role of patient-derived organoid (PDO) model in the precision medicine of advanced clear cell renal cell carcinoma (ccRCC), we retrospectively analyzed the clinical data of seven cases of ccRCC diagnosed by operation and pathology in Renji Hospital from September 2021 to September 2022. The seven patients were diagnosed with advanced ccRCC with or without remote metastasis. Cytoreductive and radical nephrectomy was performed respectively. To predict the response to immunotherapy and provide personalized medicine recommendation, a PDO model based on air-liquid interface system was established from the surgical resected tumor and subsequent drug screening was performed. Hematoxylin and eosin (H&E) staining and immunohistochemistry revealed that the PDO recapitulated the histological feature of parent tumor. Immunofluorescence staining identified that CD3+ T cells, SMA+ cancer associated fibroblasts, and CD31+ endothelial cells were preserved in PDO models. Fluorescence activated cell sorter (FACS) revealed an evidently increased ratio of CD8+/CD4+ T cells and apoptotic tumor cells in PDO treated with toripalimab than those treated with IgG4. The results showed that toripalimab is able to rescue the excessive death of CD8+ T cells by critically reversing the immune exhaustion state of ccRCC in PDO model. This research validated that PDO is a promising and faithful preclinical model for prediction of immunotherapy response in patients with ccRCC.

Purpose To develop an immunotype-based prognostic model for predicting the overall survival (OS) of patients with clear cell renal carcinoma (ccRCC). We explored novel immunotypes of patients with ccRCC, particularly those associated with overall survival. A risk-metastasis model was constructed by integrating the immunotypes with immune genes and used to test the accuracy of the immunotype model. Patients and Methods Patient cohort data were obtained from The Cancer Genome Atlas (TCGA) database, Gene Expression Omnibus (GEO) database, Renji database, and Surveillance, Epidemiology, and End Results (SEER) database. We employed the R software to select 3 immune cells and construct an immunotype-based prediction model. Immune genes selected using random Forest Algorithm were validated by immunohistochemistry (IHC). The H&L risk-metastasis model was constructed to assess the accuracy of the immunotype model through Multivariate COX regression analysis. Result Patients with ccRCC were categorized into immunotype H subgroup and immunotype L subgroup based on the overall survival rates. The immunotypes were found to be the independent prognostic index for ccRCC prognosis. As such, we constructed a new immunotypes-based SSIGN model. Three immune genes associated with difference between immunotype H and L were identified. An H&L risk-metastasis model was constructed to evaluate the accuracy of the immunotype model. Compared to the W-Risk-metastasis model which did not incorporate immunotypes, the H&L risk-metastasis model was more precise in predicting the survival of ccRCC patients. Conclusion The established immunotype model can effectively predict the survival of ccRCC patients. Except for mast cells, T cells and macrophages are positively associated with the overall survival of patients. The three immune genes identified, herein, can predict the survival rate of ccRCC patients, and expression of these immune genes is strongly linked to poor survival. The new SSIGN model provides an accurate tool for predicting the survival of ccRCC patients. H&L risk-metastasis model can effectively predict the risk of tumor metastasis.

Electrospinning using biocompatible polymer scaffolds, seeded with or without stem cells, is considered a promising technique for producing fibrous scaffolds with therapeutic possibilities for ischemic heart disease. However, no optimal scaffolds for treating ischemic heart disease have been identified thus far. In the present study, it was evaluated whether electrospun silk fibroin (SF)-blended poly(L-lactic acid-co-ε-caprolactone) [P(LLA-CL)] scaffolds that were seeded with cluster of differentiation 117 (c-kit)+ bone marrow (BM) cells may serve a protective role in cardiac remodeling following myocardial infarction (MI). Mechanical characteristics and cytocompatibility were compared between SF/P(LLA-CL) and P(LLA-CL) electrospun nanofibrous scaffolds in vitro. It was observed that MI led to a significant increase of the c-kit+ BM cell subpopulation in mice. Magnetic activated cell sorting was performed to harvest the c-kit+ cell population from the BM of mice following MI. c-kit+ BM cells were seeded on SF/P(LLA-CL) and P(LLA-CL) electrospun nanofibrous scaffolds. Results indicated that SF/P(LLA-CL) electrospun nanofibrous scaffolds were superior to P(LLA-CL) electrospun nanofibrous scaffolds in improving c-kit+ BM cell proliferation. Additionally, compared with pure SF/P(LLA-CL) electrospun nanofibrous scaffolds, SF/P(LLA-CL) scaffolds seeded with c-kit+ BM cells resulted in lower levels of MI markers and reduced infarct size, leading to greater global heart function improvement in vivo. The findings of the present study indicated that SF/P(LLA-CL) electrospun nanofibrous scaffolds seeded with c-kit+ BM cells exert a protective effect against MI and may be a promising approach for cardiac regeneration after ischemic heart disease.

346 Background: Alterations in DNA damage repair (DDR) genes are associated with human tumorigenesis and may be as potential biomarkers for vascular endothelial growth factor (VEGF) tyrosine kinase inhibitor (TKI) therapy in renal cell carcinoma. However, biologic significance and relevance to TKI targeted therapy in metastatic RCC are unknown. Methods: Genomic data and treatment outcomes were retrospectively collected for patients with metastatic RCC. Tumor and germline DNA were subject to targeted next generation sequencing across 642 genes of interest, including 60 DDR genes. Patients were dichotomized according to underlying DDR gene alteration into (1) DDR gene alterations present (Mut DDR); (2) wildtype (WT) DDR gene alterations present (WT DDR). Association between DDR status and therapeutic benefit was investigated separately for and vascular endothelial growth factor (VEGF) tyrosine kinase inhibitor (TKI) therapy. Results: Mut DDR were detected in 17/40 patients (42.5%). The most frequently DDR altered genes were TP53. For patients with TKI treatment, Mut DDR status was associated with superior progression free survival (log-rank p = 0.048), but not with superior overall survival (log-rank p = 0.39); after adjusting for International Metastatic Renal Cell Carcinoma Database Consortium risks and extent of prior therapy, the HR for Mut DDR was 2.68 (95% CI: 0.96–7.46; p = 0.059). Conclusions: DDR alterations are recurrent genomic events in patients with advanced RCC and were mostly clonal in this cohort. Dysfunction events in these genes may affect outcome with TKI therapy in adanced RCC, and these hypothesis-generating results deserve further study.