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Preferential apical infection of Caco-2 intestinal cell monolayers by SARS-CoV-2 is associated with damage to cellular barrier integrity: Implications for the pathophysiology of COVID-19

SARS-CoV-2 can infect different organs, including the intestine. In an in vitro model of Caco-2 intestinal cell line, we previously found that SARS-CoV-2 modulates the ACE2 receptor expression and affects the expression of molecules involved in intercellular junctions. To further explore the possibility that the intestinal epithelium can serve as an alternative infection route for SARS-CoV-2, we used a model of polarized monolayers of Caco-2 cells (or co-cultures of two intestinal cell lines: Caco-2 and HT29) grown on the polycarbonate membrane of Transwell inserts, inoculated with the virus either in the upper or lower chamber of culture to determine the tropism of the virus for the apical or basolateral pole of these cells. In both polarized Caco-2 cell monolayers and co-culture Caco-2/HT29 cell monolayer, apical SARS-CoV-2 inoculation was found to be much more effective in establishing infection than basolateral inoculation. In addition, apical SARS-CoV-2 infection triggers monolayer degeneration, as shown by histological examination, measurement of trans-epithelial electrical resistance, and cell adhesion molecule expression. During apical infection, the infectious viruses reach the lower chamber, suggesting either a transcytosis mechanism from the apical side to the basolateral side of cells, a paracellular trafficking of the virus after damage to intercellular junctions in the epithelial barrier, or both. Taken together, these data indicate a preferential tropism of SARS-CoV-2 for the apical pole of the human intestinal tract and suggest that infection via the intestinal lumen leads to a systemic infection.

A robust deep learning framework for multiclass skin cancer classification

Evaluating survival post-7 mo PSA response, stratified by response, disease extent, and intermittent vs. continuous ADT in mHSPC: The S9346 trial.

268 Background: A decline in prostate-specific antigen (PSA) following androgen deprivation therapy (ADT) is a well-established prognostic marker in metastatic hormone-sensitive prostate cancer (mHSPC). While S9346 trial found intermittent ADT (IAD) to be not non-inferior to continuous ADT (CAD) in mHSPC, de-intensification strategies are being considered for patients with complete PSA response. We hypothesize that IAD has different treatment effect in men with partial and complete PSA response. Methods: In the phase 3 S9346 trial men with mHSPC were randomized to IAD and CAD, if they had PSA ≤ 4.0 ng/ml after 7 months (PSA-7mo) of treatment with ADT and bicalutamide. We evaluated the association of complete PSA-7mo response (CR, PSA ≤ 0.2 ng/ml) and partial PSA -7mo response (PR, PSA 0.3-4 ng/ml) with subsequent overall survival (OS) in patients with mHSPC treated with either IAD or CAD in S9346 trial using Cox regression. We evaluated association of the disease extent and OS, using Cox regression: minimal disease defined as metastasis confined to the spine, pelvic bones, or lymph nodes; and extensive disease defined as metastasis involving ribs, long bones, or visceral organs. Results: The analysis included 1523 patients from the S9346 trial. In the IAD arm (n=763), 483 men (63%) achieved CR and 280 (37%) had PR. In the CAD arm (n=760), 473 men (62%) achieved CR and 287 (38%) had PR. A PSA-7mo CR was associated with significantly improved OS compared to PR (HR 0.57, 95% CI 0.51-0.65, p<0.0001). Extensive disease was associated with worse OS compared to minimal disease (HR 1.3, CI 1.15-1.47, p<0.0001). However, the relative treatment effect of IAD remained consistent across these subsets of patients: IAD vs CAD in subset of patients with CR HR 1.15, 95% CI 0.96-1.39; and in patients with PR HR 1.14, 95% CI 0.98-1.34. There was no statistically significant pairwise interaction between PSA response, extent of disease, and IAD vs. CAD in the multivariate OS Cox analysis (all p≥ 0.20). Conclusions: This study reaffirms the prognostic significance of a PSA-7mo CR and disease extent. IAD had the same relative treatment effect for those with a PSA CR vs. PR. These findings indicate that IAD is not an optimal therapy even in patients with good prognostic baseline clinical features and on treatment response. Clinical trial information: NCT00002651 . Multivariate Model (n=1523) Hazard Ratio 95% CI p-value CR vs. PR PSA-7mo response 0.57 (0.51, 0.65) <0.0001 Extensive vs. Minimal Disease 1.30 (1.15, 1.47) <0.0001 IAD vs. CAD 1.14 (1.02, 1.29) 0.027 IAD Arm Subset Median OS (95% CI) In months CAD Arm Subset Median OS (95% CI) In months PSA CR, ext. disease (n=228) 71 (61,83) PSA CR, ext. disease (n=193) 71 (58, 82) PSA CR, min. disease (n=255) 77 (66, 95) PSA CR, min. disease (n=280) 92 (85, 103) PSA PR, ext. disease (n=147) 43 (35, 53) PSA PR, ext. disease (n=167) 44 (36, 50) PSA PR, min disease (n=133) 43 (37, 55) PSA PR, min disease (n=120) 60 (43, 76)

Adrenal androgens and overall survival (OS) in men with castration-resistant prostate cancer (CRPC) treated with enzalutamide without (ENZ) or with abiraterone and prednisone (ENZ/AAP) (Alliance).

230 Background: DHEA, testosterone (T) and androstenedione (AD) are adrenal androgens that can drive CRPC. HSD3B1 encodes for 3β-hydroxysteroid dehydrogenase-1 (3βHSD1) and has 2 common missense-encoding germline variants: the more active adrenal-permissive allele and the less active adrenal-restrictive allele that are associated with greater and lesser non-gonadal androgen biosynthesis from DHEA. A single adrenal-permissive HSD3B1 (1245C) allele is sufficient to confer more rapid development of CRPC and homozygous inheritance (about 7-10% of men) is the most common monogenic link to prostate cancer mortality. Alliance A031201 (NCT01949337) is a phase 3 trial of ENZ versus ENZ/AAP for metastatic CRPC. This analysis combines clinical outcomes associated with adrenal androgens and HSD3B1 as the gene encoding the enzyme that uses dehydroepiandrosterone (DHEA). Methods: Germline DNA was genotyped for HSD3B1 in 929 men from A031201 (469 ENZ and 460 ENZ/AAP). Baseline DHEA, T and AD were assessed in 922 men (464 ENZ and 458 ENZ/AAP) using mass spectrometry. The Cox proportional hazards model was used to determine the prognostic significance of DHEA, T and AD categorized as tertiles in predicting OS. Results: In the combined treatment arms, median (95% CI) OS for men in low, medium and high DHEA tertiles was 27 (25, 31), 36 (33, 42) and 39 (37, 47) months (Table). In the ENZ arm, men with inheritance of 0, 1 and 2 adrenal-permissive alleles had median OS of 34 (30, 39), 34 (29, 39) and 28 (23, 37) months. Conclusions: DHEA is the most informative adrenal androgen. Men with the lowest tertile for this substrate of 3βHSD1 have the shortest OS. These data are consistent with prior analyses demonstrating poor prognosis for men with low adrenal androgens. Outcomes for men homozygous for the adrenal-permissive HSD3B1 allele treated with ENZ are consistent with prior retrospective studies. Overall survival and adrenal androgens. Serum Androgen Median OS in months (95% CI) Multivariable Hazard Ratio (95% CI) DHEA 1 st Tertile (Low) DHEA 2 nd Tertile (Medium) DHEA 3 rd Tertile (High) 27 (25, 31)36 (33, 42)39 (37, 47) AC vs. AA 0.97 (0.82,1.14)CC vs. AA 1.08 (0.81,1.44)DHEA: Med vs High 1.17 (0.97,1.42)DHEA: Low vs. High 1.65 (1.37,2.00) AD 1 st Tertile (Low) AD 2 nd Tertile (Medium) AD 3 rd Tertile (High) 32 (27, 36)36 (34,43)34 (31, 39) AC vs. AA 0.96 (0.82,1.13)CC vs. AA 1.08 (0.81, 1.44)AD: Med. vs. High 0.89 (0.73,1.07)AD: Low vs. High 1.16 (0.97,1.40) T 1 st Tertile (Low) T 2 nd Terile (Medium) T 3 rd Tertile (High) 28 (26, 35)37 (34, 41)36 (32, 41) AC vs. AA 0.95 (0.81,1.12)CC vs. AA 1.07(0.80,1.43)T: Med vs. High 1.01 (0.83,1.22)T: Low vs. High 1.28 (1.06,1.54)

EPIC-B: Phase II trial of cemiplimab as first-line treatment in advanced penile carcinoma.

9 Background: Patients with locally advanced or metastatic penile squamous cell carcinoma (la/mPC) have a poor prognosis with very limited therapeutic treatment options. Standard of Care (SoC) treatment remains platinum-based combination chemotherapy with modest outcomes. Some patients are ineligible for chemotherapy and have very limited options for management. PDL1 is upregulated in 40–60% of PC, making a case for immunotherapy as a treatment for la/mPC. We describe the results from the EPIC-B trial, evaluating the efficacy and safety of cemiplimab as first line treatment in la/mPC. Methods: EPIC-B is a National Cancer Research Network badged single arm, multi-center phase II trial, in treatment naïve la/mPC. Patients received cemiplimab 350mg IV D1 every 3 weeks (Q3W) up to a total of 34 cycles. The primary end point was investigator assessed Clinical Benefit Rate (CBR) at 12 weeks using RECIST 1.1. The study was designed as an A’Hern (2001) study α=0.05 + power (1-β)=0.8 assuming 5% meeting the primary end point is a poor treatment (p0=0.05) and 25% is a good treatment (p1=0.25). Assuming a 10% drop out rate, 18 patients were recruited to test this hypothesis. Results: From November 2021, 18 patients from 11 UK sites were enrolled onto EPIC-B over 29 months. Median age was 72 years (range 44-88). Patients with ECOG 0-2 were allowed onto the trial, the majority having ECOG 1 or 2 (0=5%, 1=56% and 2=39%). 83% had metastatic disease (1 bone 5.6%, 1 liver 5.6%, 6 lung 33.3%). Median number of cycles was 4 (range 1-32) and median follow-up was 5.5 (IQR 2.3-8.1) months. At 12 weeks CBR was 38.9% (95%CI 20.3%, 61.4%) and Objective Response Rate (ORR) was 16.6% (95%CI 5.8%, 39.2%) with 3 partial response (PR) and 4 stable disease (SD). Over the full course of treatment 27.7% of patients showed a response to treatment including 1 complete response (1 CR, 4 PR) and 4 patients had SD as their best response. Median Progression Free Survival (PFS) was calculated to be 2.3 (95%CI 1.0, 3.8) months and median Overall Survival (OS) is currently estimated at 6.8 (95%CI 3.7, 9.8) months. For adverse events (AEs) of any grade, 31% were judged related to cemiplimab and for grade 3 AEs 26% were related, most common being infection (no G4 AEs were recorded). There were 2 grade 5 AEs, (cardiorespiratory event not related and toxic epidermal necrolysis related to treatment). 4 patients discontinued treatment due to toxicity, 2 related to cemiplimab (11%). Conclusions: Single agent cemiplimab as a first line treatment for la/mPC in the EPIC-B trial demonstrates efficacy with a generally manageable toxicity profile. This study adds to the evidence that single agent cemiplimab is a viable treatment for la/mPC patients for whom chemotherapy is not an option. Clinical trial information: 95561634.

Safety and efficacy of intravesical bacillus Calmette-Guerin instillation for superficial recurrence following bladder-sparing therapy of muscle invasive bladder cancer.

787 Background: Patients treated with bladder-preserving therapy for muscle-invasive bladder cancer are at risk of developing recurrent non-muscle-invasive bladder cancer. This study aims to evaluate the efficacy and adverse events of postoperative Bacillus Calmette-Guerin (BCG) instillation in patients with superficial recurrence following bladder-preserving therapy for muscle-invasive bladder cancer. Methods: We retrospectively analyzed 120 patients diagnosed with non-muscle-invasive bladder cancer who underwent transurethral resection followed by BCG instillation. The 19 patients with prior muscle-invasive bladder cancer were categorized as the NMIBC-M group, while the remaining 101 patients formed the NMIBC group. All patients completed a six-cycle BCG course. Results: Both groups showed no significant differences in baseline characteristics, except for the number of BCG instillations (15.5 vs. 9, p = 0.010). Overall survival, recurrence-free survival, metastasis, bladder preservation, and adverse event profiles were comparable between groups. However, patients in the group with prior muscle-invasive bladder cancer reported lower rates of urinary retention (28.7% vs. 0.0%, p = 0.017) and fever (30.7% vs. 5.3%, p = 0.044). Conclusions: Postoperative BCG instillation in patients with superficial recurrence following bladder-preserving therapy for muscle-invasive bladder cancer showed comparable efficacy and side effects to those without muscle-invasive bladder cancer history.

Enhanced stability and mobility of aligned In2O3 nanofiber field-effect transistors with Y2O3 passivation

Field-effect transistors (FETs) based on indium oxide (In2O3) nanofibers demonstrate significant potential for applications in next-generation electronic devices. However, In2O3 nanofiber FETs typically exhibit deteriorated electrical performance and bias stability due to the disordered arrangement of nanofibers and a high concentration of oxygen vacancy defects. In this study, In2O3 nanofibers were prepared by electrospinning, and the effects of nanofiber orientation and Y2O3 passivation on FET electrical performance were systematically investigated. The results indicate that after Y2O3 passivation, the aligned In2O3 nanofiber FETs exhibit enhanced electrical performance and superior positive bias stress and negative bias illumination stress stability. The Y2O3 passivation layer effectively prevents the penetration of external O2 and H2O molecules, while the diffusion of Y3+ into the back channel reduces oxygen vacancies, thereby improving device stability. When Al2O3 was employed as the dielectric layer, the electrical performance of aligned In2O3 nanofiber FET with Y2O3 passivation was further optimized, achieving a mobility of 18.2 cm2/V s and a subthreshold swing of 85 mV/dec. Meanwhile, the FET exhibits excellent environmental stability after 60 days of atmospheric exposure. This work provides a strategy for fabricating nanofiber-based FETs with high mobility and stability.

Bronchoalveolar lavage single-cell transcriptomics reveals immune dysregulations driving COVID-19 severity

The continuous threats posed by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the virus that causes COVID-19, including the emergence of potentially more infectious and deadly variants, necessitate ongoing studies to uncover novel and detailed mechanisms driving disease severity. Using single-cell transcriptomics, we conducted a secondary data analysis of bronchoalveolar lavage fluid (BALF) from COVID-19 patients of varying severities and healthy controls to comprehensively examine immune responses. We observed significant immune cell alterations correlating with disease severity. In severe cases, macrophages showed upregulation of pro-inflammatory genes TNFα and IL1β, contributing to severe inflammation and tissue damage. Neutrophils exhibited increased activation, marked by S100A8, CXCL8, and IL1β expression, with extended viability and reduced phagocytosis. Genes such as MCL1 and HIF1α supported extended viability, while MSR1 and MRC1 indicated reduced phagocytosis. Enhanced formation of neutrophil extracellular traps (NETs) and reduced clearance, indicated by NET-associated markers, were linked to thrombo-inflammation and organ damage. Both macrophages and neutrophils in severe cases showed impaired efferocytosis, indicated by decreased expression of MSR1 and TREM2 in macrophages and downregulation of FCGR3B in neutrophils, leading to the accumulation of apoptotic cells and exacerbating inflammation. Severe cases were characterized by M1 macrophages with high TNFα and IL1β, while milder cases had M2 macrophages with elevated PPARγ. Dendritic cells (DCs) in severe cases exhibited reduced proportions and attenuated expression of MHC class I genes (HLA-A, HLA-B, HLA-C) and co-stimulatory molecules (CD80, CD86), alongside increased cytochrome c expression, indicating impaired antigen presentation and enhanced apoptosis. NK and T cells in severe cases demonstrated altered receptor and gene expression, with increased activation markers IFNγ and ISG15, suggesting a paradoxical state of activation and exhaustion. This analysis highlights the critical role of dysregulated neutrophil, macrophage, dendritic cell, NK, and T cell responses in severe COVID-19, identifying potential therapeutic targets and providing novel insights into the disease.

Model sensitivity limits attribution of greenhouse gas emissions to polar bear demographic rates

Abstract Greenhouse gas emissions continue to increase and negatively affect sea ice conditions that polar bears rely on. It is therefore important to better understand how specific emissions levels affect polar bear demography. A recent study proposed a framework to address this issue, but sensitivity to decisions rules of the approach may limit its utility. We tested how sensitive the approach is to decisions rules related to sea ice concentration, choice of subpopulation boundaries, and modeling choices for bears in the Chukchi Sea and Southern Beaufort Sea subpopulations. We found that the number of ice-free days, number of fasting days, and when 10% of reproductive females exhibited recruitment failure varied considerably depending on equally-valid decisions rules versus those used in the existing study. Whereas the previous study suggested that both subpopulations surpassed the critical number of ice-free days that negatively affect recruitment, we found this threshold was never reached by the Southern Beaufort Sea subpopulation and only once for the Chukchi Sea subpopulation for the decision rules we considered. Our results suggest that the previously published approach is too sensitive to modeling assumptions and choice of decision rules to accurately evaluate the impacts of GHG emissions on polar bear demographic rates.

Association of myeloid-derived suppressor cell (MDSC) dynamics with clinical response to nivolumab in metastatic clear cell renal carcinoma patients (mRCC): Results from the I-RENE Meet-URO 8 study.

586 Background: Immune checkpoint blockade is a standard-of-care treatment for mRCC patients, but the immune mechanisms driving clinical benefit remain underexplored. In the I-RENE trial (NCT04891055), we conducted a comprehensive evaluation of myeloid-derived suppressor cell (MDSC) and lymphoid dynamics and their impact on the efficacy of nivolumab. Methods: The I-RENE study is a prospective, translational, real-world multicenter trial involving mRCC patients treated with nivolumab after failure of previous VEGFR-targeted therapies. Sixty patients were enrolled between December 2018 and August 2022. Peripheral blood (PB) samples were collected at baseline and at 2, 4, and 12 weeks, as well as at disease progression. PBMCs were analyzed by high-resolution flow cytometry (profiling 144 lymphoid and myeloid cell subsets), and plasma was assessed using multiplex analysis (69 soluble factors). CD14+ monocytes from PB and tumor tissue were subjected to RNA sequencing. Results: Significant immune modulations in PB were detectable as early as 2 weeks into treatment across all patients, regardless of clinical response, and persisted throughout the 3-month observation period. These changes included substantial alterations in the profiles of immune cells and plasma soluble factors. Responders to nivolumab exhibited a marked reduction in monocyte subsets associated with immune suppression, such as CD14+HLA-DR- cells, CD14+PD-L1+ cells, and intermediate monocytes. In contrast, non-responders showed a progressive increase in suppressive monocytes and a concomitant decrease in CD14+ cells involved in anti-tumor immunity, including non-classical monocytes and CD14+ cells expressing HLA-DR and CX3CR1. Lymphoid compartment analysis revealed that responders experienced an increase in CD8+ T cells and CD4+ effector memory T cells, along with a reduction in CD38+ T cells. Conversely, non-responders showed significant upregulation of senescent CD8+ T cells (KLRG1+CD28-CD57+) and CD38+ T cells. All progressing patients showed a notable resurgence of suppressive myeloid cells (HLA-DR-, PD-L1+), confirmed by an enriched MDSC gene signature, as well as an increase in PB CD38+ and senescent T cells. Notably, the detrimental role of myeloid-driven immune suppression in non-responders was further confirmed by an enriched MDSC gene signature in matched tumor samples. Conclusions: The pervasive and detrimental impact of myeloid cells committed to immune suppression is evident even before radiological progression in mRCC patients and persists despite immunotherapy. These findings highlight the need for early identification of immune-suppressive infiltrates and the development of strategies to overcome or reprogram these cells, both at the systemic and tumor level, following diagnosis.

Frequent overexpression of NECTIN-4 in CNS metastases from urothelial carcinoma.

804 Background: The combination of enfortumab vedotin plus pembrolizumab (EV/P) has become the new standard of care for advanced or metastatic urothelial carcinoma (mUC) due to its high efficacy. However, patients with active central nervous system (CNS) metastases were excluded from the EV-302 trial. More recently, NECTIN-4 amplifications have been identified as genomic predictors of EV response and long-term survival in patients with mUC. Additionally, early evidence suggests that EV may cross the blood-brain barrier and be effective in patients with CNS metastases. Therefore, we aimed to evaluate NECTIN-4 expression and amplification in patients with mUC involving CNS metastases. Methods: Immunohistochemical staining for NECTIN-4, with H-score evaluation, was performed in both non-CNS mUC (n=152) and CNS mUC patient cohorts (n=17). In addition, we developed a NECTIN-4-specific fluorescence in situ hybridization (FISH) assay to correlate NECTIN-4 copy number variations (CNVs) with membranous NECTIN-4 protein expression in CNS mUC. Results: Membranous NECTIN-4 protein expression was more abundant in CNS mUC (n=12/17; with H-score ≥ 100; median H-score 160; interquartile range [IQR] 250-85) than in non-CNS mUC samples (n=58/152; median H-score 40; IQR 0-140, p<0.001). Amplification data are currently being generated and will be presented at the conference. Conclusions: The substantial expression of NECTIN-4 in CNS metastases from UC, along with the results of the EV-302 trial and the identification of NECTIN-4 amplifications as genomic predictors of EV response and long-term survival in patients with mUC, supports the potential use of EV/P in treating patients with CNS mUC.

Clinical characteristics and treatment patterns of patients with muscle invasive bladder cancer: A real-world cohort study.

698 Background: Due to the evolving treatment landscape for muscle-invasive bladder cancer (MIBC), a better understanding of current treatment patterns among real-world patients is needed. This study reported demographic and clinical characteristics and examined the real-world treatment patterns of patients with MIBC in the US. Methods: This non-interventional, retrospective cohort study extracted data from the Inovalon Insights Payer-Sourced dataset, a claims database, from January 1, 2020–December 31, 2021. Patients aged ≥18 years with MIBC who were treated with cystectomy with or without neoadjuvant systemic treatment were followed from index date (first day of cystectomy or neoadjuvant systemic treatment) until the end of the study, loss of follow-up, diagnosis of metastasis, or end of enrollment in their medical or pharmacy health plan, whichever occurred first. Patients with metastasis from tumor types such as solid tumors, leukemia, lymphoma and, myeloma, with secondary malignancies, or participating in clinical trials by the index date were excluded. Results were reported for patients who received neoadjuvant and/or adjuvant treatment by therapy type. Results: Among 332 eligible patients, the mean (standard deviation [SD]) age was 64.3 (10.2) years, and 245 (73.8%) patients were male. Prior transurethral resection of a bladder tumor was reported for 236 (71.1%) patients. Common comorbidities were hypertension (198 [59.6%]), urinary tract infection (94 [28.3%]), diabetes without chronic complications (88 [26.5%]), and chronic pulmonary disease (80 [24.1%]). The mean (SD) age-adjusted Charlson Comorbidity Index was 6.3 (2.3). A total of 137 (41.3%) patients underwent cystectomy alone, while 195 (58.7%) patients received neoadjuvant systemic therapy followed by cystectomy. A total of 27 (8.1%) patients received both neoadjuvant and adjuvant therapies. In the 195 patients receiving neoadjuvant systemic therapies, 153 (78.5%) received chemotherapy with cisplatin; most commonly cisplatin + gemcitabine (76 [39.0%]) followed by cisplatin + doxorubicin + methotrexate + vinblastine (56 [28.7%]). Chemotherapy without cisplatin (41 [21.0%]) and immunotherapy (1 [0.5%]) were also given as neoadjuvant systemic therapies. Adjuvant therapies were received by 39 (11.7%) patients; of which, 20 (51.3%) received immunotherapy, 15 (38.5%) received chemotherapy with cisplatin, and 4 (10.3%) received chemotherapy without cisplatin. Cisplatin + gemcitabine (10 [25.6%]) and nivolumab monotherapy (10 [25.6%]) were the most common adjuvant treatments. Conclusions: Patients with MIBC had a substantial comorbidity burden. Many patients did not receive neoadjuvant or adjuvant treatment, indicating a persistent unmet need in this patient population for alternative therapeutic regimens. Further evaluation of treatment patterns is needed as the landscape evolves.

Neoadjuvant androgen deprivation therapy with enzalutamide plus the glucocorticoid receptor modulator relacorilant versus placebo for patients with high-risk localized prostate cancer.

TPS436 Background: Despite definitive local treatment, high risk localized prostate cancer (PC) patients (pts) have a 36-46% risk of biochemical recurrence (1). Neoadjuvant (NAJ) treatment with androgen receptor signaling inhibition (ARSI) improves pathologic outcome at the time of prostatectomy (2), and 3-year biochemical recurrence-free survival (bRFS) was strongly tied to pathologic complete response (pCR) plus minimal residual disease (MRD). One way to improve on this strategy is to target potential mechanisms of resistance to ARSI. A study of localized high-risk prostate cancer treated with NAJ ARSI plus androgen deprivation therapy (ADT) showed enrichment of glucocorticoid receptor (GR) expression in residual tumors (3). Inhibiting GR signaling decreases resistance to the ARSI enzalutamide (4). We previously demonstrated the safety of the combination of the selective GR modulator (SGRM) relacorilant plus enzalutamide (5). We have thus designed a study evaluating the efficacy of this combination given neoadjuvantly. Methods: This phase 2 study is a placebo-controlled randomized trial of 6 months of NAJ ADT and enzalutamide plus relacorilant/placebo (2:1). The primary objective of this study is response, measured by pCR and MRD at radical prostatectomy (RP). Eligible patients include those with localized histologically confirmed prostatic adenocarcinoma classified as high risk or very high-risk per NCCN guidelines. Enlarged lymph nodes below the iliac bifurcation are allowed. Following randomization pts will be treated with LHRH agonist plus enzalutamide with or without relacoriliant and undergo RP 1 month later. The total sample size is 90 patients with an interim analysis for futility conducted after 45 pts undergo RP. A chi-square test will be used to compare the proportion of patients achieving pCR/MRD. Assuming a true CR/MRD rate of 15% in the control group, 90 patients total would yield a power of 80% with a hypothesized CR/MRD of 32% in the relacorilant group, based on a one-sided test at the alpha=0.15 significance level. As secondary endpoints, we will evaluate radiographic response within the prostate with multiparametric MRI (mpMRI) imaging and the 3-year bRFS and MFS rates in both arms. Exploratory endpoints include demonstrating correlation between enhanced mpMRI imaging and pathologic response, as well showing decreased nuclear hormone receptor-driven proliferative gene expression in viable PC due to combined NAJ GR antagonism and ARSI compared to ARSI alone. The study is currently open and is seeking additional sites. 1. Falgario U, JAMA Netw Open 2023. 2. McKay R, J Urol 2021. 3. Efstathiou E, Eur Urol 2019. 4. Isikbay M, Horm Cancer 2014. 5. Desai, CCR , 2024. Clinical trial information: NCT05726292 .

Anti-irradiation reinforcement in NiFe/oxide composite structure by electronic reconstruction and structural stabilization for efficient magnetoresistive sensor in aerospace/radiotherapy applications

The construction of irradiation-tolerant anisotropic magnetoresistance (AMR) sensors is crucial for weak-field detection in scenarios of aerospace and radiotherapy. Presently, the utilization of the NiFe/oxide composite structure was considered to be an effective scheme to optimize the spin-dependent transport property; however, it exhibited poor anti-irradiation ability due to the crystal instability of oxide. Here, a strategy was proposed to break through the limitation based on the electronic reconstruction and structural stabilization. By introducing an oxygen-affinitive Hf intercalation into the Ta/MgO/NiFe/MgO/Ta multilayer, the electron coordination was modified to tune the 3d orbital occupancy of Fe, apparently boosting the s-d electron scattering and spin-related transport property. Meanwhile, the irradiation stability of electronic and crystal structures was effectively improved due to the emergence of the Hf–O–Mg bond with high dissociation energy. Therefore, we constructed a highly reliable AMR sensor with both the ultrahigh sensitivity of 3.1 mV/V/Oe and excellent irradiation-tolerant ability capable of resisting the γ-ray irradiation of 1000 Gy. These results not only build an important basis for the sensor application in the irradiation environment but also provide a possible idea for the anti-irradiation design in spintronic devices.

Pathology and parasitology of free-ranging coyotes from Tennessee and South Carolina

Coyotes are exposed to many parasites and pathogens of veterinary and zoonotic concern. To assess the prevalence of the diseases caused by these microbes, we opportunistically obtained coyote samples from a variety of sources including a GPS collaring study, rabies testing facilities, wildlife resources agents, and road-side mortalities. We performed necropsies, serological testing, fecal flotations, and molecular analyses on coyotes from Tennessee and South Carolina. Dirofilaria immitis (heartworm) infected 46% (41/89) of coyotes and was associated with eosinophilic alveolitis and arteritis. Paragonimus kellicotti, a zoonotic lung fluke, was found in 24% (17/71) of Tennessee coyotes, including one coyote with extrapulmonary infection affecting the liver and lymph nodes. Trichinella spp., a zoonotic nematode, was present in 17% (12/71) of Tennessee coyotes but was not associated with muscular inflammation. Sarcoptes scabiei, the causative agent of sarcoptic mange, was detected in one Tennessee coyote. Most coyotes (86% [90/105]) were seropositive for Toxoplasma gondii, while 8.5% (9/106) were seropositive for Trypanosoma cruzi, an emerging zoonotic, vector-borne parasite. This study demonstrated that coyotes are commonly exposed to numerous parasites and pathogens that affect people and pets and are excellent sentinels for these diseases.

Vericiguat prevents high glucose-mediated impaired vascular smooth muscle cGMP production and vasorelaxation

Risk stratification using the Decipher 22-gene genomic classifier (GC) and digital pathology artificial intelligence (DPAI) in nearly 10,000 localized prostate cancer patients.

408 Background: DPAI models have recently demonstrated the potential to improve risk stratification beyond routine clinical and pathologic variables. However, it is not known whether integrating DPAI information will enhance the prognostic accuracy of validated gene expression tests. In this study, the prognostic performance of novel DPAI algorithms were assessed in context to the genomic classifier (GC). Methods: A prospectively collected cohort of 9,874 patients with localized prostate cancer was retrieved from the Veracyte GRID registry (NCT02609269). Scans at 40X magnification were obtained for 17,701 H&E slides using an Aperio GT450 scanner (Leica). An open-source whole-slide pathology foundation model (GigaPath) was used to encode whole slide image (WSI) patches into digital pathology image features (DPIF). Attention-based multiple instance learning approach was used to develop separate models to predict distant metastasis (DM) in the biopsy and radical prostatectomy (RP) training subsets. WSI, baseline clinical variables, and GC scores were linked using tokenization (Datavant) to real-world data (Clarivate). The primary endpoint of this study was DM. Adjusted Hazard ratio (aHR) from multivariable Cox regression (MVA) modeling and 5-year area-under curve (AUC) estimates were used to compare models using DPIF. Results: Median follow-up for the training cohort (n=6,705; 239 DM events) and validation (n=3,169; 110 DM events) cohorts were 6.5 years. In the biopsy validation cohort of 999 patients, the GC predicted DM with an AUC of 0.80 (95% CI, 0.70-0.90), which exceeded NCCN risk group alone (AUC 0.68, 0.56-0.80) and DPAI (AUC 0.76, 0.66-0.86). MVA including age, NCCN, GC score, and DPAI, only showed GC (aHR 1.23 [1.03, 1.47]) and DPAI (aHR 1.22 [1.04, 1.43]) to be significantly associated with risk of DM (aHR per 10%, both p<0.05). However, an integrated model with clinicopathologic features, DPAI, and GC did not improve discrimination (AUC 0.80, 0.70-0.90). In the RP validation cohort of 1,492 patients, GC predicted DM with an AUC of 0.81 (95% CI, 0.73-0.88). An integrated model with clinicopathologic features, DPAI, and GC improved the AUC to 0.84 (0.77- 0.91). In MVA, only RP GC (aHR 1.27 [1.13, 1.43]) and DPAI (aHR 1.44 [1.23, 1.69]) were significant predictors for DM (both p<0.001). Conclusions: To our knowledge, this is the largest study assessing the prognostic value of adding DPAI to improve performance above and beyond a clinical-genomic model. The results from this study suggest that the combination of these data sources may further improve prognostication, and use of both DPAI and genomics negated information from routine clinical variables. Ongoing efforts in larger cohorts are underway to identify optimal scenarios in which GC and DPAI information enhance clinical utility for decision making.

Trends in kidney cancer: Exploring the impact of sex and age on stage of disease, and prognosis during the past three decades in Denmark—A DaRenCa study.

485 Background: Over the past 30 years, renal cell carcinoma (RCC) management has undergone significant advancements, driven by increased diagnosis through imaging scans and the development of more effective systemic therapies for metastatic disease. Understanding the impact of early detection and evolving treatment strategies on patient outcomes is crucial for guiding future advancements in RCC care. This nationwide registry-based cohort study investigates how the number of RCC cases, stage at diagnosis, and prognosis have changed during the past 30 years in Denmark, and how these are associated with sex and age. Methods: A population-based cohort was established, including all Danish patients aged 18 and older diagnosed with RCC from 1992 to 2021, excluding individuals with a prior cancer history. Patients were identified and followed from the date of diagnosis until death or end of follow-up using nationwide register data. Comorbidities prior to RCC diagnosis were assessed using nationwide hospital admittance records. Results: A total of 17,423 RCC patients were diagnosed during the past 30 years in Denmark. The male-to-female ratio changed significantly from 0.59:0.41 in 1992-1996 to 0.72:0.28 in 2017-2021 (P < 0.001). The age distribution of patients at diagnosis remained relatively constant (ranging from 65-67 years). The number of RCC cases increased from 2,244 in 1992-1996 to 3,947 in 2017-2021. We observed stage migration over time, with Stage I cases increasing from 30% in 2002-2006 to 55% in 2017-2021, and Stage IV cases decreasing from 45% in 2002-2006 to 20% in 2017-2021, while the absolute number of metastatic cases remained relatively unchanged. The median survival of patients with metastatic disease steadily increased by 314% from 4.1 months in 1992-1996 to 12.9 months in 2017-2021. Additionally, married patients showed better survival than divorced and unmarried patients. Conclusions: During the past 30 years, the number of RCC cases in Denmark has increased, primarily driven by early-stage tumors, while the number of metastatic cases has remained stable. Additionally, the proportion of female patients has decreased across all stages of RCC during the observation period. Overall survival has significantly improved over time, highlighting the impact of early detection and treatment advancements. Despite only studying Danish patients, this study serves as a reference for how patient dynamics have evolved with the advancements in the diagnosis and treatment of RCC.

Targeting TROP2 in aggressive variant and neuroendocrine prostate cancer.

171 Background: Aggressive variant prostate cancer (AVPC) including neuroendocrine prostate cancer (NEPC) is characterized by rapid, androgen-independent tumor growth. Standard therapies often show limited efficacy. Alternative treatment strategies are therefore urgently required. Here, we present preclinical and first clinical results on the potential impact of TROP2 targeting in AVPC/NEPC. Methods: TROP2 expression was assessed in AVPC/NEPC cell lines, circulating tumor cells (CTCs) and metastatic biopsies. Efficacy of the TROP2 targeted antibody drug conjugate (ADC) Sacituzumab Govitecan (SG) was evaluated in prostate cancer (PC) cell lines with high and low/- TROP2 expression. TROP2+ AVPC/NEPC patients with exhausted standard therapy options are currently undergoing treatment with SG in an individual approach. Results: TROP2 expression was detected at varying levels in PC cell lines, with increased expression in docetaxel-resistant cells. CTCs of AVPC and NEPC patients were TROP2+ in 74.0% (37/50) and 48.6% (17/35), respectively, while the vast majority of biopsies from both cohorts expressed TROP2. SG was highly effective in TROP2+ cells in vitro , regardless of their degree of resistance to standard therapies while reduced activity was seen in TROP2 low/- cells after short-term exposure. Remarkably, SG exhibited a pronounced antiproliferative effect and/or apoptosis induction in TROP2-low LNCaP and PC3 cells, when co-cultured with TROP2+ DU145-DR or PC3-DR cells, indicating a bystander effect. The antiproliferative activity of SG was attributed to an increased G2/M cell cycle arrest. Proteomic analyses suggested suppression of cell cycle related processes and proliferation as well as signaling pathways involved in neuronal dedifferentiation and cell stemness. First clinical results of heavily pretreated TROP2+ AVPC/NEPC patients (n=9) revealed partial remission (PR) or stable disease in 44% and 33% at radiological follow-up, with a disease control rate of 78% and 44% at 3 and 6 months, respectively. Of note, in a NEPC patient with sudden, rapid progression following initial PR, loss of TROP2 expression on CTCs was detected. In addition, preclinical data suggested AKT activation and upregulation of p-glycoprotein as further possible resistance mechanisms. Conclusions: TROP2-directed therapy with SG demonstrates promising preclinical and clinical activity in AVPC and NEPC. A clinical trial evaluating a TROP2-directed ADC in AVPC is in preparation.

Observational study of tislelizumab (Tisle) combined with pazopanib in reducing venous tumor thrombus in clear cell renal cell carcinoma with venous invasion.

481 Background: Renal cell carcinoma with venous tumor thrombus (RCC-VTT) presents significant surgical challenges, often requiring complex interventions. Neoadjuvant therapies that can reduce primary tumor size and venous tumor thrombus may simplify surgical procedures and decrease associated risks. Methods: The data of pts with RCC-VTT who underwent Tisle combined with Pazopanib neoadjuvant therapy at the First Affiliated Hospital of Xinjiang Medical University from Nov. 2021 to Jan. 2024 were analyzed. Inclusion criteria were as follows: (1) age ≥ 18 years; (2) TNM stage T3-4N0-1M0-1; (3) Eastern Cooperative Oncology Group (ECOG) performance status ≤ 1; (4) a life expectancy of at least 3 months; and (5) adequate bone marrow, kidney, liver function, and normal blood test results. Exclusion criteria were as follows: (1) prior treatment with other antitumor drugs; (2) recent history of cardiac or vascular events or other conditions precluding surgical treatment; (3) participation in other clinical trials within the past 3 months; (4) lack of legal capacity or limited legal capacity; and (5) contraindications, allergies, or adverse reactions to pazopanib and tislelizumab. Eligible Pts received Tisle 200 mg IV. in day 1(D1), every 21 days for 4 cycles; Pazopanib was administered orally at a dose of 800 mg once daily for 12 weeks prior to the scheduled surgery. The primary endpoint of the study was the percentage change in the length of the VTT and the percentage reduction in the longest tumor diameter. Results: Nine patients with a median age of 58 years were treated. The median reduction in tumor diameter was 31.0% (range: 3.0%–48.7%), and the median reduction in VTT length was 29.2% (range: -153.1% to 100.0%). Treatment led to decreased surgical complexity in 88.9% of patients. The regimen was well-tolerated, with manageable adverse events primarily including vomiting, itching, and weight loss. No metastasis developed during the study period, though one patient required expedited surgery due to an increase in thrombus size. Conclusions: The combination of tislelizumab and pazopanib as neoadjuvant therapy effectively reduces the size of tumors and associated venous tumor thrombi in patients with RCC-VTT, facilitating less complex surgical interventions and potentially leading to better postoperative outcomes.