Adverse events in CA-3 vs vehicle treated mice

Inhibition of α2 reduces invasiveness of PNE and PAC cells

INTRODUCTION:Male infertility is increasingly recognized as a systemic condition associated with impaired long-term health, yet clinically applicable frameworks linking reproductive dysfunction to comorbidity burden remain limited. Here, we investigated whether a composite immuno-metabolic signature could capture comorbidity burden and reduced reproductive capacity in men with primary male factor infertility (MFI). METHODS AND MATERIALS:In a cross-sectional cohort of 2953 men, metabolic and inflammatory variables were systematically screened for directional and independent associations with comorbidity burden, defined by the Charlson Comorbidity Index (CCI) ≥1, and impaired reproductive capacity, assessed by total motile sperm count (TMSC) <5 million. Three variables-LDL cholesterol, waist circumference, and C-reactive protein (CRP)-met selection criteria and were integrated into an unweighted standardized score. RESULTS:Three variables met selection criteria: LDL cholesterol, waist circumference, and C-reactive protein. Higher immuno-metabolic scores were observed in men with comorbidity burden (0.20 ± 0.73 vs. -0.02 ± 0.62, p < 0.001) and in those with TMSC <5M (0.13 ± 0.64 vs. -0.07 ± 0.62, p < 0.001). Each unit increase in the score was independently associated with comorbidity burden (OR 1.50, 95% CI 1.23-1.82, p < 0.001) and TMSC <5M (OR 1.67, 95% CI 1.48-1.89, p < 0.001). Predicted probabilities rose across the score range from 2% to 40% for comorbidity and from 12% to 75% for reduced reproductive capacity. CONCLUSION:These findings identify a biologically coherent immuno-metabolic signature linking spermatogenic impairment with systemic comorbidity in men with primary MFI and support a model in which male infertility reflects broader immuno-metabolic vulnerability. Prospective external validation is warranted before clinical implementation.

Divergence of the proteomes of ECMs obtained from TRAMP and WT prostates

Polarized light analysis of ECM linearization in human prostate cancer specimen

Differentially expressed matrisome genes discriminate human NEPC and CRPC-Adeno

Gene signatures used for transcriptomic analysis

Upregulation of integrin α2 is characteristic of PNE and not PAC cells

The combination of systemic immune checkpoint inhibitors with standard of care intravesical Bacillus Calmette-Guerin (BCG) therapy for non-muscle invasive bladder cancer (NMIBC) has shown potential in enhancing BCG therapeutic effects. However, challenges such as disease recurrence, progression, and immune-related toxicities remain a significant hurdle. Furthermore, the mechanism of action of BCG involves broad, non-specific immune activation. While this recruits bystander CD8+ and NK cells, it also triggers the undesired expansion of Tregs, which may ultimately hamper therapeutic efficacy. We aimed to explore the feasibility of a targeted approach designed to overcome intravesical tumor resistance and selectively enhance the expansion and effector functions of CD8+ TILs. We tested intravesical administration of BCG with murinized PD1-IL2v, a fusion protein that simultaneously targets PD-1 and IL-2Rβγ in cis on the same cell, in a preclinical mouse model of NMIBC. Both BCG monotherapy and co-treatment with a murinized PD1-IL2v improved animal survival. Notably, the intravesical combination of BCG and murinized PD1-IL2v significantly increased the number of CD8+ TILs with polyfunctional cytotoxic effector phenotype and avoided Treg expansion in contrast to BCG monotherapy. In addition, in patient-derived tumors, we observed a higher frequency of CD8⁺ TILs (18

PNE cells translocate YAP into the nucleus when cocultured with PAC cells

Abstract The emergence of the neuroendocrine phenotype in castration-resistant prostate cancer (CRPC) is associated with poor patient prognosis. Castration-induced death of fully differentiated, androgen-sensitive prostate cancer cells might foster interactions among rare androgen-independent, poorly differentiated cancer cells and the extracellular matrix (ECM) that promotes the development of neuroendocrine prostate cancer (NEPC). In this study, we investigated physical and molecular interactions between poorly differentiated prostate cancer cells with exocrine (PAC) or neuroendocrine features (PNE), which recapitulated preexisting human CRPC-like cells, and decellularized prostate ECM. Without androgens, PAC cells and prostate cancer–derived ECM promoted in vitro invasiveness of PNE cells by inducing integrin α2 upregulation and YAP activation, indicating a cell-to-cell and cell-to-matrix contact-driven process. Inhibition of RANK/RANKL and NF-κB prevented integrin α2 upregulation in PNE cells, and integrin α2β1 and YAP inhibition also reduced PNE invasiveness. Microenvironment-conditioned PNE cells showed YAP-dependent metastatic behavior in vivo, and YAP inhibition suppressed the development of NEPC and metastasis in castration-naïve mice and of neuroendocrine CRPC in transgenic mice with prostate cancer. Importantly, YAP inhibitors also restrained the growth of human CRPC organoids. These findings unveil mechanisms of NEPC development and implicate the integrin α2–YAP axis as a therapeutic target in patients with prostate cancer receiving androgen deprivation therapy. Significance: Targeting signaling pathways activated by interactions between poorly differentiated neuroendocrine and exocrine prostate cancer cells and the surrounding ECM suppresses NEPC development and metastasis.

Signature of pre-existing CRPC-like cells segregates NEPC from adenocarcinoma in mice and humans

CONTEXT:Antimüllerian hormone (AMH) is produced by Sertoli cells and may serve as a biomarker of spermatogenesis, yet the literature lacks comprehensive comparative analyses across male fertility conditions. OBJECTIVE:This work aimed to discover how serum AMH levels differ among men with varying fertility statuses, and the relationship between AMH, spermatogenesis, and functional testicular reserve. METHODS:A cross-sectional study was conducted of 1085 White European, non-Finnish men with confirmed fertility, primary infertility, or nonazoospermic infertility. Men with confirmed fertility (n = 116), primary infertility (n = 791), and nonobstructive azoospermia (NOA) (n = 178) underwent comprehensive hormonal and semen analyses per World Health Organization 2010 criteria. Kruskal-Wallis and chi-square tests were used for group comparisons. Correlations were assessed using Spearman rank correlation. Multivariable linear regression models identified factors associated with AMH levels and sperm concentration. RESULTS:Serum AMH levels were significantly lower in men with NOA compared to fertile controls and NOA infertile men. AMH correlated negatively with age and follicle-stimulating hormone (FSH), and positively with testicular volume and sperm concentration. After adjustment, AMH showed no independent association with sperm concentration. AMH levels were significantly lower in NOA vs fertile men and primary infertility 3.8 (1.6-7.2) vs 5.1 (3.6-7.0) vs 4.9 (3.0-7.8) ng/mL; P less than .001. AMH negatively correlated with age and FSH, positively with testicular volume and sperm concentration. Age, FSH, and testicular volume were independently associated with AMH; FSH and testicular volume, not AMH, were independently associated with sperm concentration. However, the cross-sectional design limits causality, and the study faced limited ethnic generalizability. CONCLUSION:AMH reflects Sertoli cell function and testicular status rather than directly influencing spermatogenesis, potentially serving as a complementary male fertility biomarker.

Efficacy of CA-3 treatment in mouse models of PC

CA-3 inhibits NEPC and not WT organoids

α2 upregulation requires cell-to-cell contact and NF-kB signaling

CONTEXT:Non-obstructive azoospermia (NOA) is the most severe form of male infertility, affecting 1% of all men, with a clinical picture characterized by no sperm production, hyalinization of the basal membrane of the seminiferous tubules, primary hypogonadism, and earlier onset of age-related comorbidities compared with fertile men. NOA is also characterized by etiologic heterogeneity and the non-genetic form has higher incidence of testicular germ cell cancer (TGCC) compared to the forms with genetic abnormalities. OBJECTIVE:We aimed to establish molecular pathways in the testicular somatic cells that are either shared or specific for non-genetic and genetic forms of NOA, such as complete androgen insensitivity syndrome (CAIS) and Klinefelter syndrome (KS). METHODS:We performed single-cell RNA sequencing of the testicular somatic cells of an individual with CAIS, and data integration with published scRNA-seq datasets of testis with normal spermatogenesis, NOA, KS, and germinal testicular cancer. Detailed clinical data of the CAIS patient, testosterone and estradiol levels in age-matched men (120 fertile, 155 infertile, 116 NOA, 18 KS, and 343 with TGCC) were analyzed. RESULTS:In all conditions, Leydig cells are immature and senescent, but those of NOA associated with primary hypogonadism depict the highest expression of transcripts associated with the seminoma microenvironment, including estrogen-responsive genes. An oncological transcriptional signature in the Leydig cells has been confirmed at the systemic levels by showing a prognostic role of the decreasing testosterone/estradiol ratio for TGCC in men with non-genetic NOA. CONCLUSION:This study offers molecular insights into the prediction of TGCC in persons with NOA and eligibility for the use of aromatase inhibitors.

Identified proteins in the mouse ECM