The quest for a non-hormonal male contraceptive pill for men still exists. Serine protease 37 (PRSS37) is a sperm-specific protein that when ablated in mice renders them sterile. In this study we sought to examine the molecular sequelae of PRSS37 loss to better understand its molecular function, and to determine whether human PRSS37 could rescue the sterility phenotype of knockout (KO) mice, allowing for a more appropriate model for drug molecule testing. To this end, we used CRISPR-EZ to create mice lacking the entire coding region of Prss37, used pronuclear injection to create transgenic mice expressing human PRSS37, intercrossed these lines to generate humanized mice, and performed LC-MS/MS of KO and control tissues to identify proteomic perturbances that could attribute a molecular function to PRSS37. We found that our newly generated Prss37 KO mouse line is sterile, our human transgene rescues the sterility phenotype of KO mice, and our proteomics data not only yields novel insight into the proteome as it evolves along the male reproductive tract, but also demonstrates the proteins significantly influenced by PRSS37 loss. In summary, we report vast biological insight including insight into PRSS37 function and the generation of a novel tool for contraceptive evaluation.
Abstract Background: Only one quarter of patients with muscle invasive urothelial bladder cancer (MIBC) gain approximately 5% improvement in 5-year overall survival from cisplatin-based neoadjuvant chemotherapy (NAC). For patients with residual invasive cancer post radical cystectomy there is no standard of care and high mortality. Previous TCGA projects focused on pre-NAC MIBC genomic and transcriptomic alterations and identified 5 molecular subtypes with differential risk and response. For this study we hypothesized that comprehensive proteomic and phosphoproteomic profiling of MIBC prior to NAC will further define mechanisms responsible for chemotherapy resistance, and identify specific and actionable targeted therapies for patients with NAC-resistant tumors. Methods: OCT-embedded and flash frozen tissue samples from 143 eligible patients were processed and tested for proteomics quality control (QC). Samples containing >45% tumor content, less than 10% muscle content and >2,000 protein identifications in a single-shot quality control assay were selected for deep-scale proteomic and phosphoproteomic profiling. A final cohort of 60 samples (52 pre-treatment and 8 patient-matched post-treatment tumors) were multiplexed using tandem mass tags (TMT-11), fractionated by basic reverse phase chromatography and analyzed by liquid chromatography and mass spectrometry (LC-MS). Results: Over 12,000 proteins were identified in total, and 8,353 proteins identified in all samples, including 425 kinases and 77 targets of FDA approved cancer therapies. Principal component analysis identified two distinct resistant clusters and one sensitive cluster. The samples were clustered based on the 5 TCGA subtypes which resulted in the sub stratification of the resistant clusters into two resistance-enriched basal-squamous clusters (BS1 and BS2 respectively), one infiltrated-luminal cluster (L1), one sensitive luminal cluster (L2) and one intermediary cluster (L3) (with 86% samples resistant). UV response, Epithelial to Mesenchymal transition and Myogenesis were significantly elevated in both L1 and L3 (resistant) relative to L2 (sensitive). Similar, these pathways are significantly altered in L1 relative to L3. EGFR, CDK6, ITPKC and CSNK1 were elevated in the B4 subtype, all of which are potentially druggable targets. Matched samples and phosphoproteomics data will be presented. Conclusion: Surrounding non-tumor tissue can obfuscate true tumor signatures in MIBC, cohort selection through stringent pathologic QC allows proteomic profiling to identify tumor features that correlate with NAC resistance. This dataset provides proof of principle that actionable targets and subtype distinctions can be identified through discovery proteomics and further analysis in NAC treated cohorts are justified. Citation Format: Matthew V. Holt, Meggie N. Young, Karoline Kremers, Alexander Saltzman, Antrix Jain, Mei Leng, Hugo Villanueva, Lacey E. Dobrolecki, Beom-Jun Kim, Meenakshi Anurag, Matthew J. Ellis, Anna Malovannaya, Seth P. Lerner. Proteomic profiling of muscle invasive bladder cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3922.
Abstract To assess the impact of inoculation of inactivated corona virus disease 2019 (COVID-19) vaccine on the outcome of frozen-thawed embryo transfer (FET). From January 2022 to November 2023, patients aged 20 ~ 45 years old undergoing FET at the Reproductive Medicine Center of a tertiary teaching hospital were prospectively enrolled. The patients were divided into vaccinated group (n = 458) and unvaccinated group (n = 530) based on the inoculation of inactivated COVID-19 vaccine before FET. Vaccinated group was further divided into three subgroups based on the dose (single dose, n = 55; double dose, n = 292; triple dose, n = 111) or interval from the first inoculation to FET (< 3 months, n = 51; 3 ~ 6 months, n = 101; > 6 months, n = 306). The primary outcome was live birth rate (LBR). The LBR (43.87% vs. 40.57%) was not significantly different between the vaccinated and unvaccinated groups, and so were embryo implantation rate (IR), clinical pregnancy rate (CPR), the gestational age at delivery and birth weight (P > 0.05). IR was significantly decreased with the shorter interval of vaccination (28.57% vs. 32.02% vs.45.24%, P = 0.007), while LBR and CPR were not significantly different (P > 0.05). For the dose subgroups of vaccination, IR, LBR and CPR were not significantly different (P > 0.05). Inoculation of inactivated COVID-19 vaccine did not affect the outcome of FET. Clinical Trial Registration Number: ChiCTR2200055597 (Chinese Clinical Trial Registry), January 14, 2022. (https://www.chictr.org.cn/bin/project/edit?pid=148312)
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