Primary cilia are increasingly recognized as regulators of cellular signaling and plasticity. Here, we examined their distribution and potential relevance in neuroendocrine (NE) prostate cancer. While typically absent in localized hormone-sensitive prostate tumor cells, we detected primary cilia in neuroendocrine-like cells both in vitro and in castration-resistant prostate cancer (CRPC) samples. In vivo, cilia were consistently observed in CRPC tumor cells exhibiting FDG-PET positivity and NE features, supporting an association between ciliogenesis, metabolic reprogramming, and disease progression. These aggressive tumors also displayed reduced mitochondrial activity, consistent with a shift away from oxidative metabolism. Building on our work in ccRCC, we identified a GLI1⁺/IFT20⁺ or GLI1⁺/IFT80⁺ signature enriched in ciliated, NE-prone subpopulations. In vitro, YAP1 inhibition alone did not induce ciliogenesis, whereas cytoskeletal remodeling with jasplakinolide restored cilium assembly and enabled partial NE transdifferentiation. Single-cell RNA-seq analyses further showed enrichment of ciliogenesis-related genes within NE clusters in CRPC. Together, these observations support a model in which primary cilia are closely associated with NE identity and metabolic adaptation, rather than serving solely as passive markers, and suggest a structural-metabolic axis that may represent a source of biomarkers and therapeutic vulnerabilities.

Worldwide, prostate cancer (PCa) ranks second in terms of incidence and eighth in terms of mortality. While most cancers remain silent after initial treatment, some tumors recur. At present, we are unable to predict which patients are at risk of recurrence. In order to determine this risk of recurrence as early as at the biopsy stage, and to enable better therapeutic management of patients, it is essential to identify new biomarkers. In this study, we have demonstrated that Cysteine Dioxygenase CDO1 could be a predictive marker in PCa progression. Transcriptomic analysis showed that CDO1 expression is significantly reduced in patients who have relapsed and lower CDO1 expression is associated with poorer survival outcomes. In PCa, CDO1 expression could be regulated by methylation and androgen signaling pathway. Furthermore, inhibition of CDO1 expression in VCaP prostate cancer cells led to increased cell migration and non-adherent growth. Finally, transcriptomic analysis of these cells with inhibited CDO1 expression demonstrates the complex role of CDO1 and its possible involvement in the mechanisms regulating endoplasmic reticulum stress and protein unfolding. Altogether, these results describe the loss of CDO1 as a marker of aggressiveness in PCa. Furthermore, the loss of CDO1 is thought to be responsible for tumor progression in vitro by acting on multiple signaling pathways.

Background: Advancing chimeric antigen receptor (CAR) T cell therapy for solid tumors remains a major challenge in cancer immunotherapy. Prostate cancer (PCa), particularly in its aggressive forms, may be a suitable target for CAR-T therapy given the range of associated tumor antigens. However, due to the high plasticity and heterogeneity of aggressive PCa and the complexity of the tumor environment, there is a need to broaden the repertoire of targetable antigens and deepen our understanding of CAR-T behavior in stressed microenvironmental conditions. Growing evidence supports mesothelin as a promising cancer-associated marker and a compelling target for CAR-T cell approaches in solid tumors. Objectives and Methods: Here, we employed gene expression datasets to investigate mesothelin expression in both primary and metastatic PCa tumors. Additionally, we evaluated mesothelin expression across various preclinical PCa models and assessed the therapeutic efficacy of second-generation mesothelin-targeted CAR-T (meso-CAR-T) cells under both normoxic and hypoxic conditions, with hypoxia as a representative tumor-associated stress condition. Results: Our results revealed a significant enrichment of mesothelin in 3–10% of metastatic prostate tumors, contrasting with its minimal expression in primary tumors. In line with these findings, we observed increased mesothelin expression in an aggressive variant of the 22Rv1 cell line, which displayed an epithelial–mesenchymal plasticity (EMP) phenotype. Meso-CAR-T cells demonstrated potent cytotoxicity and remarkable selectivity toward these carcinoma cells under both severe hypoxia (1% O2) or normoxia (21% O2), highlighting their ability to withstand metabolic stress within the tumor microenvironment. Conclusions: Our study underscores the potential of meso-CAR-T cells as a promising strategy for targeting specific subtypes of metastatic prostate cancer.

Background: Molecular understanding of muscle -invasive (MIBC) and non-muscleinvasive (NMIBC) bladder cancer is currently based primarily on transcriptomic and genomic analyses. Objective: To conduct proteogenomic analyses to gain insights into bladder cancer (BC) heterogeneity and identify underlying processes specific to tumor subgroups and therapeutic outcomes. Design, setting, and participants: Proteomic data were obtained for 40 MIBC and 23 NMIBC cases for which transcriptomic and genomic data were already available. Four BC -derived cell lines harboring FGFR3 alterations were tested with interventions. Intervention: Recombinant tumor necrosis factor -related apoptosis-inducing ligand (TRAIL), second mitochondrial-derived activator of caspases mimetic (birinapant), pan-FGFR inhibitor (erdafitinib), and FGFR3 knockdown.

In early 2020, the novel pathogenic severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in Wuhan, China, and rapidly propagated worldwide causing a global health emergency. SARS-CoV-2 binds to the angiotensin-converting enzyme 2 (ACE2) protein for cell entry, followed by proteolytic cleavage of the Spike (S) protein by the transmembrane serine protease 2 (TMPRSS2), allowing fusion of the viral and cellular membranes. Interestingly, TMPRSS2 is a key regulator in prostate cancer (PCa) progression which is regulated by androgen receptor (AR) signaling. Our hypothesis is that the AR signaling may regulate the expression of TMPRSS2 in human respiratory cells and thus influence the membrane fusion entry pathway of SARS-CoV-2. We show here that TMPRSS2 and AR are expressed in Calu-3 lung cells. In this cell line, TMPRSS2 expression is regulated by androgens. Finally, pre-treatment with anti-androgen drugs such as apalutamide significantly reduced SARS-CoV-2 entry and infection in Calu-3 lung cells but also in primary human nasal epithelial cells. Altogether, these data provide strong evidence to support the use of apalutamide as a treatment option for the PCa population vulnerable to severe COVID-19.

Polymersomes are an exciting modality for drug delivery due to their structural similarity to biological cells and their ability to encapsulate both hydrophilic and hydrophobic drugs. In this regard, the current work aimed to develop multifunctional polymersomes, integrating dye (with hydrophobic Nile red and hydrophilic sulfo-cyanine5-NHS ester as model drugs) encapsulation, stimulus responsiveness, and surface-ligand modifications. Polymersomes constituting poly(N-2-hydroxypropylmethacrylamide)-b-poly(N-(2-(methylthio)ethyl)acrylamide) (PHPMAm-b-PMTEAM) are prepared by aqueous dispersion RAFT-mediated polymerization-induced self-assembly (PISA). The hydrophilic block lengths have an effect on the obtained morphologies, with short chain P(HPMAm)16 affording spheres and long chain P(HPMAm)43 yielding vesicles. This further induces different responses to H2O2, with spheres fragmenting and vesicles aggregating. Folic acid (FA) is successfully conjugated to the P(HPMAm)43, which self-assembles into FA-functionalized P(HPMAm)43-b-P(MTEAM)300 polymersomes. The FA-functionalized P(HPMAm)43-b-P(MTEAM)300 polymersomes entrap both hydrophobic Nile red (NR) and hydrophilic Cy5 dye. The NR-loaded FA-linked polymersomes exhibit a controlled release of the encapsulated NR dye when exposed to 10 mM H2O2. All the polymersomes formed are stable in human plasma and well-tolerated in MCF-7 breast cancer cells. These preliminary results demonstrate that, with simple and scalable chemistry, PISA offers access to different shapes and opens up the possibility of the one-pot synthesis of multicompartmental and responsive polymersomes.

Supplementary Table 1 from Class III β-Tubulin Expression Predicts Prostate Tumor Aggressiveness and Patient Response to Docetaxel-Based Chemotherapy

Background Neuroendocrine prostate cancer (NEPC) is a multi-resistant variant of prostate cancer (PCa) that has become a major challenge in clinics. Understanding the neuroendocrine differentiation (NED) process at the molecular level is therefore critical to define therapeutic strategies that can prevent multi-drug resistance. Methods Using RNA expression profiling and immunohistochemistry, we have identified and characterised a gene expression signature associated with the emergence of NED in a large PCa cohort, including 169 hormone-naïve PCa (HNPC) and 48 castration-resistance PCa (CRPC) patients. In vitro and preclinical in vivo NED models were used to explore the cellular mechanism and to characterise the effects of castration on PCa progression. Results We show for the first time that Neuropilin-1 (NRP1) is a key component of NED in PCa cells. NRP1 is upregulated in response to androgen deprivation therapies (ADT) and elicits cell survival through induction of the PKC pathway. Downmodulation of either NRP1 protein expression or PKC activation suppresses NED, prevents tumour evolution toward castration resistance and increases the efficacy of docetaxel-based chemotherapy in preclinical models in vivo. Conclusions This study reveals the NRP1/PKC axis as a promising therapeutic target for the prevention of neuroendocrine castration-resistant variants of PCa and indicates NRP1 as an early transitional biomarker.

Abstract Long noncoding (lnc)RNAs modulate gene expression alongside presenting unexpected source of neoantigens. Despite their immense interest, their ability to be transferred and control adjacent cells is unknown. Extracellular Vesicles (EVs) offer a protective environment for nucleic acids, with pro and antitumourigenic functions by controlling the immune response. In contrast to extracellular nonvesicular RNA, few studies have addressed the full RNA content within human fluids’ EVs and have compared them with their tissue of origin. Here, we performed Total RNA‐Sequencing on six Formalin‐Fixed‐Paraffin‐Embedded (FFPE) prostate cancer (PCa) tumour tissues and their paired urinary (u)EVs to provide the first whole transcriptome comparison from the same patients. UEVs contain simplified transcriptome with intron‐free cytoplasmic transcripts and enriched lnc/circular (circ)RNAs, strikingly common to an independent 20 patients’ urinary cohort. Our full cellular and EVs transcriptome comparison within three PCa cell lines identified a set of overlapping 14 uEV‐circRNAs characterized as essential for prostate cell proliferation in vitro and 28 uEV‐lncRNAs belonging to the cancer‐related lncRNA census (CLC2). In addition, we found 15 uEV‐lncRNAs, predicted to encode 768 high‐affinity neoantigens, and for which three of the encoded‐ORF produced detectable unmodified peptides by mass spectrometry. Our dual analysis of EVs‐lnc/circRNAs both in urines’ and in vitro’s EVs provides a fundamental resource for future uEV‐lnc/circRNAs phenotypic characterization involved in PCa.

AbstractSmall Extracellular Vesicles (sEVs) are 50–200 nm in diameter vesicles delimited by a lipid bilayer, formed within the endosomal network or derived from the plasma membrane. They are secreted in various biological fluids, including airway nasal mucus. The goal of this work was to understand the role of sEVs present in the mucus (mu‐sEVs) produced by human nasal epithelial cells (HNECs) in SARS‐CoV‐2 infection. We show that uninfected HNECs produce mu‐sEVs containing SARS‐CoV‐2 receptor ACE2 and activated protease TMPRSS2. mu‐sEVs cleave prefusion viral Spike proteins at the S1/S2 boundary, resulting in higher proportions of prefusion S proteins exposing their receptor binding domain in an ‘open’ conformation, thereby facilitating receptor binding at the cell surface. We show that the role of nasal mu‐sEVs is to complete prefusion Spike priming performed by intracellular furin during viral egress from infected cells. This effect is mediated by vesicular TMPRSS2 activity, rendering SARS‐CoV‐2 virions prone to entry into target cells using the ‘early’, TMPRSS2‐dependent pathway instead of the ‘late’, cathepsin‐dependent route. These results indicate that prefusion Spike priming by mu‐sEVs in the nasal cavity plays a role in viral tropism. They also show that nasal mucus does not protect from SARS‐CoV‐2 infection, but instead facilitates it.

Prostate cancer (PCa) is the second most frequent cancer and the fifth leading cause of cancer death in men worldwide. If local PCa presents a favorable prognosis, available treatments for advanced PCa display limiting benefits due to therapeutic resistances. Nucleolin (NCL) is a ubiquitous protein involved in numerous cell processes, such as ribosome biogenesis, cell cycles, or angiogenesis. NCL is overexpressed in several tumor types in which it has been proposed as a diagnostic and prognostic biomarker. In PCa, NCL has mainly been studied as a target for new therapeutic agents. Nevertheless, little data are available concerning its expression in patient tissues. Here, we investigated the expression of NCL using a new cohort from Mondor Hospital and data from published cohorts. Results were then compared with NCL expression using in vitro models. NCL was overexpressed in PCa tissues compared to the normal tissues, but no prognostic values were demonstrated. Nine genes were highly co-expressed with NCL in patient tissues and tumor prostate cell lines. Our data demonstrate that NCL is an interesting diagnostic biomarker and propose a signature of genes co-expressed with NCL.

H 2 O 2 -sensitive block copolymer nanoparticles (NPs) composed of a hydrophilic macro-chain transfer agent (macro-CTA) and a hydrophobic thioether-bearing block were prepared by polymerization-induced self-assembly (PISA) approach. PISA process first involved a chain extension of poly((poly(ethylene glycol) methyl ether methacrylate)- co -(poly(ethylene glycol) methacrylate)) macro-CTA with a H 2 O 2 -responsive N-(2-(methylthio)ethyl)acrylamide monomer (MTEAM), which self-assembled into P((PEGMA-co -PEGMAOH)- b -PMTEAM) block copolymer NPs. The polymerization kinetics indicated the evolution of particle size and morphology from spherical micelles, fused micelles, to vesicles over time. Upon incubation with 0.1 to 10 mM H 2 O 2 , spheres were fragmented due to the oxidation of the PMTEAM cores, thus transforming the hydrophobic thioethers into hydrophilic sulfoxides, and disassociating the nanocarriers. 43 % of the hydrophobic Nile Red dye was encapsulated into the spherical micelles and demonstrated a controlled release in H 2 O 2 incubation. Spherical micelles and vesicles displayed no cytotoxicity in MCF-7, DU145 and 22Rv1 cells. Both spheres and vesicles were efficiently internalized into MCF-7 cells, with spheres showing higher level of uptake than vesicles due to the smaller sizes. In summary, PISA of P((PEGMA-co -PEGMAOH)- b - PMTEAM) allowed the direct formation and loading of hydrophobic dye into spherical micelles which showed a controlled drug release in H 2 O 2 .

H2O2-sensitive block copolymer nanoparticles (NPs) composed of a hydrophilic macro-chain transfer agent (macro-CTA) and a hydrophobic thioether-bearing block were prepared by polymerization-induced self-assembly (PISA) approach. The PISA process first involved the chain extension of poly((poly(ethylene glycol) methyl ether methacrylate)-co-(poly(ethylene glycol) methacrylate)) macro-CTA with a H2O2-responsive N-(2-(methylthio)ethyl)acrylamide (MTEAM) monomer, which self-assembled into P((PEGMA-co-PEGMAOH)-b-PMTEAM) block copolymer NPs. The polymerization kinetics indicated the evolution of particle size and morphology from spherical micelles, fused micelles, to vesicles over time. Upon incubation with 0.1 to 10 mM H2O2, spheres were fragmented due to the oxidation of the PMTEAM cores, thus transforming the hydrophobic thioethers into hydrophilic sulfoxides and disassociating the nanocarriers. 43% of the hydrophobic Nile red dye was encapsulated into the spherical micelles and demonstrated a controlled release in H2O2 incubation. Spherical micelles and vesicles displayed no cytotoxicity in MCF-7, DU145, and 22Rv1 cells. Both spheres and vesicles were efficiently internalized into MCF-7 cells, with spheres showing a higher level of uptake than vesicles due to the smaller sizes. In summary, PISA of P((PEGMA-co-PEGMAOH)-b-PMTEAM) allowed the direct formation and loading of hydrophobic dye into spherical micelles, which showed a controlled drug release in H2O2.

Long noncoding (lnc)RNAs modulate gene expression alongside presenting unexpected source of neoantigens. Despite their immense interest, their ability to be transferred and control adjacent cells is unknown. Extracellular Vesicles (EVs) offer a protective environment for nucleic acids, with pro and anti-tumorigenic functions by controlling the immune response. In contrast to extracellular non-vesicular RNA, few studies have addressed the full RNA content within human fluids’ EVs and none have compared them with their tissue of origin. Here, we performed Total RNA-Sequencing on 6 Formaldehyde-Fixed-Parafilm-Embedded (FFPE) prostate cancer (PCa) tumor tissues and their paired urinary (u)EVs to provide the first whole transcriptome comparison from the same patients. UEVs contain simplified transcriptome with intron-free cytoplasmic transcripts and specific lnc/circular (circ)RNAs, strikingly common to all patients. Our full cellular and EVs transcriptome comparison within 3 common PCa cell lines identified a set of overlapping 14 uEV-circRNAs characterized as essential for prostate cell proliferation in vitro and 15 uEV-lncRNAs that we predicted to encode 768 high-affinity neoantigens. Our dual analysis of EVs-lnc/circRNAs both in urines’ and in vitro’s EVs provides a fundamental resource for future uEV-lnc/circRNAs phenotypic characterization involved in PCa.

Prostate cancer (PCa) is the second most frequent cancer and the fifth leading cause of cancer death among men worldwide. At first, advanced PCa is treated by androgen deprivation therapy with a good initial response. Nevertheless, recurrences occur, leading to Castrate-Resistance Prostate Cancer (CRPC). During the last decade, new therapies based on inhibition of the androgen receptor pathway or taxane chemotherapies have been used to treat CRPC patients leading to an increase in overall survival, but the occurrence of resistances limits their benefits. Numerous studies have demonstrated the implication of extracellular vesicles (EVs) in different cancer cellular mechanisms. Thus, the possibility to isolate and explore EVs produced by tumor cells in plasma/sera represents an important opportunity for the deciphering of those mechanisms and the discovery of biomarkers. Herein, we summarized the role of EVs in therapeutic resistance of advanced prostate cancer and their use to find biomarkers able to predict these resistances.

TMPRSS2 est une protéase cellulaire régulée par les androgènes dans les cellules prostatiques. L’entrée de SARS-CoV2 par fusion membranaire dans les cellules pulmonaires nécessite le clivage de Spike par TMPRSS2. Notre hypothèse est que le niveau des androgènes et la présence du récepteur des androgènes (RA) dans les cellules pulmonaires pourrait réguler l’expression de TMPRSS2 et influencerait l’entrée de SARS-CoV-2. Les régulations de TMPRSS2 et du RA ont été étudiées dans deux lignées cellulaires pulmonaires (A549 et Calu-3) et dans des lignées de cellules prostatiques contrôles (LNCaP ou VCaP) grâce à des expériences de privation et de supplémentation en androgènes ainsi que par utilisation d’un siRNA ciblant le RA. La régulation de l’expression de ces deux gènes a été étudiée par RT-qPCR et Western blot. L’infection de SARS-CoV2 dans les cellules Calu-3 a été étudiée en condition de blocage du RA. Les études cliniques semblent montrer que les patients atteints de cancer de la prostate sous hormonothérapie ont une incidence plus faible d’infection par le SARS-CoV2 suggérant le rôle du RA dans l’infection du virus. Nos résultats montrent que les deux lignées de cellules pulmonaires utilisées (A549 et Calu-3) expriment TMPRSS2 et le RA au niveau ARNm et protéique. Dans ces lignées, il existe également une régulation de l’expression de TMPRSS2 et du RA par les androgènes. Cet effet est connu et bien retrouvé pour les lignées prostatiques. Enfin, nous avons pu montrer qu’au niveau des cellules pulmonaires, le taux d’infection par le SARS-CoV2 était moins important lorsque la voie du RA a été au préalable bloquée. Le blocage de TMPRSS2 permettrait de limiter l’infection du virus. Nos résultats suggèrent que l’expression de TMPRSS2 est régulée par les androgènes dans des cellules pulmonaires et que la testostérone pourrait jouer un rôle dans l’infection du SARS-CoV2. Le blocage du RA serait donc une option thérapeutique envisagée pour limiter la COVID-19.

Dysregulated androgen receptor (AR) plays a crucial role in prostate cancer (PCa) development, though further factors involved in its regulation remain to be identified. Recently, paradoxical results were reported on the implication of the MEN1 gene in PCa. To dissect its role in prostate luminal cells, we generated a mouse model with inducible Men1 disruption in Nkx3.1-deficient mice in which mouse prostatic intraepithelial neoplasia (mPIN) occur. Prostate glands from mutant and control mice were analyzed pathologically and molecularly; cellular and molecular analyses were carried out in PCa cell lines after MEN1 knockdown (KD) by siRNA. Double-mutant mice developed accelerated mPIN and later displayed microinvasive adenocarcinoma. Markedly, early-stage lesions exhibited a decreased expression of AR and its target genes, accompanied by reduced CK18 and E-cadherin expression, suggesting a shift from a luminal to a dedifferentiated epithelial phenotype. Intriguingly, over 60% of menin-deficient cells expressed CD44 at a later stage. Furthermore, MEN1 KD led to the increase in CD44 expression in PC3 cells re-expressing AR. Menin bound to the proximal AR promoter and regulated AR transcription via the H3K4me3 histone mark. Interestingly, the cell proliferation of AR-dependent cells (LNCaP, 22Rv1, and VCaP), but not of AR-independent cells (DU145, PC3), responded strongly to MEN1 silencing. Finally, menin expression was found reduced in some human PCa. These findings highlight the regulation of the AR promoter by menin and the crosstalk between menin and the AR pathway. Our data could be useful for better understanding the increasingly reported AR-negative/NE-negative subtype of PCa and the mechanisms underlying its development.

According different studies and reports by live gender data tracker, more infected men than women seem to be dying from the new coronavirus, in different countries hit by the pandemic [1, 2].In France, during March 2020 we observed a more pronounced excess mortality in men than in women (+ 13%) compare to March 2019 and 2018.The excess mortality was even higher in region registering high rate of .Men can be more susceptible to viruses than women.Women generally tend to have stronger immune responses to viruses, though the reason for that is still up for debate.Genetic and hormonal differences may play a role, while environmental factors could also contribute to susceptibility to different viruses.During previous epidemics of coronaviruses, male sex was associated with worse clinical outcomes due to severe acute respiratory syndrome (SARS) in Hong Kong [4], and a higher risk of dying from Middle East respiratory syndrome (MERS) [5].As specialists of prostate cancer, we were particularly interested to learn SARS-CoV-2 uses the SARS-CoV receptor ACE2 for entry and the serine protease TMPRSS2 for S protein priming.SARS-CoV-2 may even require the combined presence of ACE-2 and TMPRSS2 to enter cells [6].TMPRSS2 is a transmembrane serine protease.

Till now, prostate cancer (PC) appears among the most critical public health concerns in men in developed as well as developing countries. Androgens operating through androgen receptor (AR) nourish the development and function of normal prostate and may pathologically contribute to PC in case of any deformation or deregulation (1). Based on this, inhibiting the production of androgens by castration or their effects by using anti- AR agents is employed as a treatment of advanced PC disease. However, in most instances, upon therapy, cancer will develop in a form called castrate-resistant prostate cancer (CRPC).