Supplementary legends Supplementary Tables 1-3 Supplementary Figures 1-8

The most common cancer in men is prostate cancer. Treatment options for prostate cancer consist of radiation therapy, active surveillance, chemotherapy, and surgical intervention. In this case, radiation therapy was used, but it can lead to adverse effects ranging from fatigue, impacted sexual function, genitourinary side effects such as dysuria or nocturia, and gastrointestinal side effects such as nausea, abdominal pain, or diarrhea. Minimizing the treatment only to areas with PET-positive disease and those with a significant risk of microscopic involvement, therefore reducing radiation exposure, can help mitigate side effects and help maintain the patient's quality of life. Here, we examine a man with prostatic adenocarcinoma. A prostate-specific membrane antigen (PSMA)-PET scan was used to determine if the seminal vesicles demonstrated any evidence of disease and if treatment of the area was necessary. A PSMA-PET scan showed invasion of the seminal vesicles; thus, the seminal vesicles were included as a target for radiation therapy. The patient tolerated the treatment with minimal side effects. This case reviews the indications for treating the whole seminal vesicle.

Renal cell carcinomas account for 2-3% of all malignancies. In non-familial syndrome patients, another kidney tumor synchronous, antecedent, or subsequent to diagnosis of RCC is relatively rare. It is very rare for the kidney tumor to be malignant and of differing pathology altogether. We herein describe a 67-year old male with bilateral synchronous masses treated with open bilateral adrenal sparing radical nephrectomy. His final pathology was interestingly found to have novel, differing aggressive cancers: a TFE3-rearranged RCC and a grade 4 WHO clear cell RCC.

Melanosis of the bladder is an exceedingly rare benign condition that can mimic more serious urothelial pathologies. Here, we analyze the clinical presentation, associated symptomatology, and follow-up of a patient presenting with melanosis and associated urinary tract infection. This patient is a 72-year-old male undergoing workup for gross hematuria, lower urinary tract symptoms, and recurrent urinary infections. We subsequently discuss two additional incidental findings of melanosis and associated urinary tract infection at our institution, and introduce novel context for this condition that currently lacks a standardized management protocol. To our knowledge, this is the first case series on melanosis vesicae.

Genetic alterations play a pivotal role in various human diseases, particularly cancer. The androgen receptor (AR) is a crucial transcription factor driving prostate cancer progression across all stages. Current AR-targeting therapies utilize competitive AR antagonists or pathway suppressors. However, therapy resistance often emerges due to AR mutations and AR splice variants, such as AR-v7. To overcome this, we developed ATC-324, an AR degrader using the innovative protein degradation technology platform AUTOphagy-TArgeting Chimera (AUTOTAC). ATC-324 was designed to comprise enzalutamide, an AR inhibitor, as a target-binding ligand and YT 6-2, a ligand of the autophagy receptor p62/SQSTM1, as an autophagy-targeting ligand. ATC-324 induces the formation of the AR/p62 complex, leading to autophagy-lysosomal degradation of AR. Importantly, ATC-324 effectively degrades AR mutants frequently detected in prostate cancer and codegrades AR-v7 as a heterodimer with full-length AR. ATC-324 reduces nuclear AR levels and downregulates the target gene expression of AR and AR-v7, leading to cytotoxicity in AR-positive prostate cancer cells. We also provide evidence of the therapeutic potential of ATC-324 in vivo as well as ex vivo bone organ culture. Moreover, ATC-324 remains potent in enzalutamide-resistant prostate cancer cells. These results demonstrate the potential of the AUTOTAC platform to target previously considered undruggable proteins and overcome certain drug resistance mechanisms. Significance: The characterization of an AUTOTAC-based degrader capable of inducing autophagic degradation of wild-type and mutated androgen receptors demonstrates the potential of this approach for targeting castration-resistant prostate cancer and overcoming drug resistance.

Abstract Prostate cancer (PCa) progression is largely driven by the androgen receptor (AR), making it a prime target for therapy. However, therapy resistance often arises due to AR mutations and splice variants, such as AR-v7. AUTOTAC (AUTOphagy-TArgeting Chimera) is a novel protein degradation platform that utilizes the autophagy-lysosomal pathway to selectively degrade disease-causing proteins. In this study, we characterize ATC-324, an AUTOTAC-based AR degrader designed for the treatment of PCa. ATC-324 comprises enzalutamide linked to YT 6-2, a small molecule activator of the autophagy receptor p62/SQSTM1. ATC-324 induces the formation of the AR/p62 complex while activating autophagic flux, leading to the autophagy-lysosomal degradation of AR. To characterize ATC-324, we employed various AR-dependent, AR-null, and enzalutamide-resistant PCa cell lines. Given that bone metastasis is a major clinical complication of castrate-resistant PCa associated with greater morbidity and mortality, we utilized the bone-in-culture array (BICA) assay, an ex vivo bones culture model, to investigate the impact of ATC-324 on PCa cell survival and growth in the bone microenvironment. We demonstrated that ATC-324 effectively degrades wild-type, mutant, and splice variants of AR. In addition, ATC-324 induces AR-dependent apoptosis and cytotoxicity. Importantly, ATC-324 maintains its potency in enzalutamide-resistant PCa cells. Utilizing the BICA assay, we showed that ATC-324 exhibits cytotoxicity against 22Rv1 bone micrometastases at even lower concentrations than in 2D cell culture. In conclusion, our study demonstrates the successful development and characterization of ATC-324 as a potent and selective degrader of various forms of AR. Furthermore, its effectiveness against enzalutamide-resistant cells indicates its capability to overcome therapy resistance. The observed cytotoxicity against bone micrometastases suggests a therapeutic potential for combating bone metastasis, a major complication of castration-resistant PCa. Our study underscores the potential of the AUTOTAC platform to selectively degrade disease-causing proteins Citation Format: Tri Pham, Tae Hyun Bae, Ki Woon Sung, Abdo Najy, Alaleh Zamiri, Hyejeong Jang, Su Ran Mun, Seongho Kim, Dongping Shi, Steven Kregel, Elizabeth Heath, Michael Cher, Yong Tae Kwon, Hyeong-Reh Kim. Development and characterization of ATC-324: An AUTOTAC-based androgen receptor degrader for prostate cancer treatment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6045.

Background: Melanosis vesicae is a rare condition characterized by the deposition of melanin within the bladder urothelium. Case presentation: We present a case of a 72-year-old male with a history of recurrent urinary retention, bladder diverticula, and concurrent Aerococcus urinary tract infection who presented with left-sided abdominal pain. Cystoscopy revealed diffuse black splotch lesions throughout the bladder and two diverticula. Histopathological examination confirmed the diagnosis of melanosis vesicae. The patient ultimately underwent an open bladder diverticulectomy. Conclusion: The potential associations between melanosis vesicae, urinary tract malignancies and concurrent conditions such as bladder diverticula and urinary infections warrant further investigation.

PURPOSE:Bladder cancer is predominantly a disease of older individuals. Concurrent chemotherapy and radiation is a bladder-sparing strategy for management of muscle-invasive bladder cancer; however, many patients are not candidates for chemotherapy due to comorbidities or impaired performance status. We conducted a study in a chemotherapy-ineligible patient population with the objectives of evaluating the safety, efficacy, and quality-of-life effect of the combination of nivolumab and radiation therapy in patients with localized/locally advanced urothelial cancer. METHODS AND MATERIALS:Eligible patients had muscle-invasive bladder cancer and were not candidates for standard chemoradiation strategy due to at least one of the following: performance status of 2, creatinine clearance ≤60 mL/min, cardiac disease, neuropathy, and intolerance to previous treatment. Creatinine clearance ≥40 mL/min, normal marrow, and liver function were required. The primary endpoint was progression-free survival at 12 months. Nivolumab was started within 3 days of radiation therapy and administered at a dose of 240 mg intravenously every 2 weeks for a maximum of 6 months. Radiation therapy was per standard of care for bladder cancer. Imaging and cystoscopy and biopsy evaluation were required at months 3, 6, and 12 and then annually until progression. RESULTS:Twenty patients were enrolled, with a median age of 78.5 years (range, 58-95 years); 80% of patients were >70 years of age, and 8 (40%) were >80 years of age. Median creatinine clearance was 52 mL/min. Nine patients (48%) were progression free at 12 months. Median progression-free survival was 11.4 months (90% CI, 7.5-23.7 months), and median overall survival was 15.6 months (90% CI, 9.1-26.1 months). CONCLUSIONS:Concurrent nivolumab and radiation therapy is tolerable but demonstrated limited efficacy in an older population with multiple comorbidities. Immune correlates demonstrated that patients with baseline programmed cell death ligand 1 combined prognostic score ≥5% had numerically longer progression-free survival.

AbstractBackgroundThe Discoidin Domain Receptor 1 (DDR1) is one of the two members of a unique family of receptor tyrosine kinase receptors that signal in response to collagen, which has been implicated in cancer progression. Here, we examined the expression of DDR1 in prostate cancer (PCa), and assessed its potential value as a prognostic marker, as a function of grade, stage and other clinicopathologic parameters.MethodsWe investigated the association between the expression level and subcellular localization of DDR1 protein and PCa aggressiveness by immunohistochemistry, using tissue microarrays (TMAs) encompassing 200 cases of PCa with various Gleason scores (GS) and pathologic stages with matched normal tissue, and a highly specific monoclonal antibody.ResultsDDR1 was found to be localized in the membrane, cytoplasm, and nuclear compartments of both normal and cancerous prostate epithelial cells. Analyses of DDR1 expression in low GS (≤ 7[3 + 4]) vs high GS (≥ 7[4 + 3]) tissues showed no differences in nuclear or cytoplasmic DDR1in either cancerous or adjacent normal tissue cores. However, relative to normal-matched tissue, the percentage of cases with higher membranous DDR1 expression was significantly lower in high vs. low GS cancers. Although nuclear localization of DDR1 was consistently detected in our tissue samples and also in cultured human PCa and normal prostate-derived cell lines, its presence in that site could not be associated with disease aggressiveness. No associations between DDR1 expression and overall survival or biochemical recurrence were found in this cohort of patients.ConclusionThe data obtained through multivariate logistic regression model analysis suggest that the level of membranous DDR1 expression status may represent a potential biomarker of utility for better determination of PCa aggressiveness.

Involvement of the adrenal gland in kidney cancer represents a unique site of metastasis with a distinct clinical course. The cases are typically resistant to immune therapy and need local therapy management. A case series of patients with adrenal metastases was reviewed to highlight the nuances of clinical course and therapy. We reviewed renal cancer carcinoma (RCC) cases with adrenal metastases at Karmanos Cancer Center, Detroit MI. Medical records were reviewed to collect relevant case information. Next-generation sequencing, tumor mutation burden testing, and programmed death ligand biomarkers were evaluated in five cases. Twelve cases were reviewed; all were males with a median age of 49.5 years. Three patients presented with adrenal metastases only and were treated with local therapy. Three received interleukin-2 (IL-2). One patient relapsed with bilateral adrenal lesions after 11 years of remission, post-IL-2 therapy. Five cases received immune checkpoint inhibitor (ICI) and one received antivascular therapy. ICI therapy was followed by ablation of residual adrenal metastases in three patients. Genomic profiling was available in five cases. All were BAP1 and PD-L1 negative.Pathogenic mutations in PBRM1, SETD2, and VHL were noted. All patients with residual adrenal metastases responded to antivascular therapies or to local ablation. One patient died 17 years after diagnosis and 11 patients are alive at a median follow-up of 9.5 years. Adrenal metastases in RCC have a distinct clinical course. They can represent a sanctuary site of relapse/residual disease following treatment with immune therapy. Management with local therapy can induce durable remissions. Systemic management with antivascular therapies also demonstrated favorable responses. Further investigation should focus on the unique clinical course and optimal management of adrenal metastases in kidney cancer.

INTRODUCTION: Herpes simplex virus (HSV) esophagitis is rarely seen in immunocompetent patients. Symptoms include odynophagia, dysphagia, fever, and/or extra-esophageal herpetic lesions. A male predominance is noted with individuals being inflicted with the illness in the third to fourth decade. Although HSV esophagitis has typical punch-out lesions on endoscopy, it is very similar in appearance to cytomegalovirus (CMV) and should be confirmed by tissue biopsy. Upper gastrointestinal bleeding is a rare complication of HSV esophagitis. Treatment of HSV esophagitis in immunocompetent patients is controversial. CASE DESCRIPTION/METHODS: A 63-year-old African American woman with past medical history of chronic obstructive pulmonary disease (COPD) presented with acute shortness of breath and productive cough despite adherence to home medications. She was admitted for the management of a COPD exacerbation and treated with intravenous methylprednisolone, inhaled steroid (budesonide), bronchodilators (albuterol-ipratropium), and doxycycline. Prompt respiratory demise required intubation and transfer to the intensive care unit. She subsequently developed coffee ground emesis with associated drop in hemoglobin. An esophagogastroduodenoscopy (EGD) was performed which revealed punched-out esophageal ulcers and a non-bleeding superficial antral ulcer. Cold-forceps biopsies were taken from esophageal ulcers, which showed typical HSV histologic changes (multinuclear cells with ground glass appearance nuclei). Diagnosis was confirmed by positive immunohistochemical staining for HSV and negative staining for CMV. She was then treated with oral valacyclovir for 10 days. Repeat EGD for percutaneous endoscopic gastrostomy placement (1 day after completion of valacyclovir therapy) demonstrated complete resolution of previously noted esophagitis. Patient did not have any more gastrointestinal bleeding during admission. DISCUSSION: HSV esophagitis, although rare, can occur in immunocompetent patients. A high index of suspicion is required as patients may present with upper gastrointestinal bleeding but without any systemic symptoms as in our case. HSV esophagitis should be considered in immunocompetent patients with COPD or asthma exacerbation who are receiving systemic steroid therapy and develop upper gastrointestinal bleeding. Valacyclovir may be used successfully in treatment of HSV esophagitis in immunocompetent patients.

Pathologic complete response is an exceptionally rare occurrence in prostate cancer, especially in the setting of poorly differentiated cancer, with high risk and poor prognostic features. Patient reviewed and signed an informed consent. The case details were collected. Patient had PSA of 52.6 ng/dl and Gleason score 5 + 5 = 10 prostate adenocarcinoma with focal signet ring cell pattern. Genomic testing revealed pathogenic p53 and SPOP mutations. The patient received androgen deprivation therapy and six cycles of docetaxel. His PSA declined to undetectable, and radical prostatectomy (RP) showed no evidence of malignancy. The patient has discontinued all therapy and continues in remission 12 months after surgery.

Objectives: To estimate the probability of downgrading to Gleason score <= 7 at radical prostatectomy for men with a prostate needle biopsy demonstrating Gleason score 8 (4 + 4). Methods: This is a retrospective review of men with Gleason score 8 (4 + 4) prostate cancer on needle biopsy who then underwent a radical prostatectomy at the Karmanos Cancer Institute or the University of Michigan. Men with any pattern 5 on the diagnostic biopsy were excluded. The objective was to estimate the proportion of patients whose tumors were downgraded to Gleason score <= 7 at radical prostatectomy and to identify clinical and biopsy parameters associated with downgrading. Results: Median age of our cohort was 63 years (IQR: 59, 67.5) and median follow-up was 15 months (IQR: 7, 37). Of the 105 men that met inclusion criteria, 59% (62/105) were downgraded to Gleason score <= 7 at radical prostatectomy. Having <= 2 cores demonstrating Gleason score 8, <= 50% maximal tumor involvement of any individual core positive for Gleason score 8, or the presence of Gleason pattern 3 (such as 3 + 4, 4 + 3, or 3 + 3) in other biopsy cores were all independently associated with downgrading in our multivariable model. Depending on the absence, presence, or combination of these 3 factors, patients had an estimated 6% to 82% probability of having their tumor downgraded at radical prostatectomy. Conclusions: Men with low volume Gleason 8 (4 + 4) and/or the presence Gleason pattern 3 on prostate needle biopsy often have their tumors downgraded at radical prostatectomy. The presence of these preoperative biopsy parameters could affect pretreatment counseling and impact patient management. Published by Elsevier Inc.

Background: Gleason grade 4 (GG4) PC has different architectural patterns with % of GG4 being an important prognosticator in patients(pts) with GS 7(3+4) PC.Recent reports indicate that CFPC may be an additional, independent adverse risk factor in this PC category.We aimed to explore the potential association of CFPC with aggressive clinicopathologic parameters in men with GS 7(3+4) PC on Bx.Design: All pts diagnosed with Bx GS 7(3+4) PC between 2005-2011 in our institution were included.PSA at diagnosis, no. of positive cores, % of core involvement, % of GG4 and presence of CFPC were documented.The presence of CFPC was correlated with post treatment parameters; stage and margin status following radical prostatectomy (RP); biochemical recurrence(BR), distant metastasis(DM), progression free survival(PFS) and cancer specific mortality(CSM) for all pts.Statistical analyses were conducted using KM and regression analysis.Results: 283 pts qualified for analyses.We classified 74%,53%,26% and 21% of GG4 as fused glands, poorly formed glands, glomeruloid and CFPC, respectively.At diagnosis, pts with CFPC had significantly higher PSA, more positive cores, higher % core involvement and higher % of GG4 (P=0.03,P=0.001,P=0.001and P=0.001,respectively).Forty percent of pts were treated with RP, 55% by radiation therapy(RT), 3% cryoablation and 2% chose active surveillance.After RP, GS was significantly upgraded for pts with CFPC(P=0.03).Median follow up was 89 (range 4-150)months.BR and DM were significantly higher in pts with CFPC compared to those without (P=0.001& P=0.001,respectively).On univariate regression analysis, CFPC was a significant predictor of BR(P=0.001),DM(P=0.003) and CSM(P=0.01).On multivariate analysis, CFPC was an independent predictor of CSM(P=0.02).When stratifying pts based on treatment, CFPC was significantly associated with BR(P=0.03) and an independent predictor of CSM(P=0.02) after RP.Following RT, BR was also significantly higher in pts with CFPC(P=0.009).On KM analysis, pts with CFPC had significantly shorter BR free survival and PFS(P=0.001 and P=0.008, respectively).PFS was also significantly shorter in pts with CFPC who underwent RP(P=0.04),while pts who received RT had shorter BRFS(P=0.001)and PFS(P=0.01).Conclusions: Our data strongly suggest that CFPC, as a variant of GG4 PC, is significantly associated with poorer clinicopathololgic features.Accounting for this variant has the potential of influencing prognostic and management considerations.

Abstract Discoidin domain receptors DDR1 and DDR2 are the only receptor tyrosine kinases that bind to and signal in response to collagen. In cancer, DDRs have been shown to play a key role in mediating the crosstalk between tumor cells and the stromal collagen matrix. Because prostate cancer (PCa) preferentially metastasizes to bone, a collagen-rich microenvironment, we set out to investigate the role of DDR1 in intraosseous growth of PC3 cells, a human PCa cell line that expresses DDR1 but not DDR2. PC3 cells were engineered to express short hairpin RNAs (shRNAs) against DDR1, or a scrambled shRNA as a control. These cells were inoculated into the tibiae of male SCID mice, and then bone response and intraosseous tumor growth evaluated by X-ray and histomorphometry. Whereas no differences were observed in bone response (osteolytic lesions), downregulation of DDR1 in PC3 cells was associated with a significant increase in intraosseous tumor growth when compared to control PC3 cells (P<0.05, Mann-Whitney test). To further evaluate the role of DDR1 in PC3 tumor growth, human DDR1a was overexpressed in PC3 cells. The engineered cells were inoculated subcutaneously into SCID mice and tumor growth was monitored over time. Tumors formed by DDR1a-overexpressing PC3 cells were significantly smaller than those obtained with control cells (P<0.005, at final time point, Mann-Whitney test). We also evaluated the expression of DDR1 in a 200-case grade/stage tissue microarray (TMA) with matched normal tissue obtained from The Prostate Cancer Biorepository Network (PCBN). Tissues were subjected to immunohistochemistry (IHC) with an antibody to human DDR1. A predominant membranous DDR1 immunostaining was observed in most epithelial prostate cells, with sporadic cytoplasmic or nuclear immunoreactivity. We found no differences in DDR1 expression between normal and benign prostate glands and low-grade PCa (Gleason score ≤7 [3+4]). However, a significantly lower DDR1 expression was observed in high-grade PCa (Gleason score ≥7 [4+3]) when compared to low-grade PCa (P=0.002, Fisher's exact test). These results suggest that reduced DDR1 expression in PCa is associated with a more aggressive disease. Collectively, these data highlight an unreported role of DDR1 in PCa progression whereby DDR1 may elicit a tumor-suppressive activity, as shown by its ability to reduce intraosseous and subcutaneous growth of PC3 cell xenografts. These data also suggest a differential role for DDRs in PCa bone metastasis whereby DDR1, as opposed to DDR2 (as reported previously), restrains intraosseous PCa growth. Because DDRs are being considered as potential novel therapeutic targets in cancer, defining their precise contributions in PCa progression is of critical importance for the development of effective therapies in this cancer type. Citation Format: R. Daniel Bonfil, Anjum Sohail, Semir Vranić, Daniel S. Oliveira, Dongping Shi, Wei Chen, Hyejeong Jang, Allen D. Saliganan, Benjamin D. Wasinski, Hyeong-Reh C. Kim, Rafael A. Fridman. Discoidin domain receptor 1 (DDR1): A potential suppressor of prostate cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1070.

Introduction: Histologic transformation from non-small cell lung cancer (NSCLC) to small cell lung cancer (SCLC) is a well-recognized mechanism of resistance in EGFR-mutant adenocarcinoma upon treatment with TKIs, but rarely reported with programmed death1 (PD-1) inhibitors. We report two cases of potential transformation during treatment with PD-1 inhibitors. Case presentations: Case 1, a 65-year-old man was diagnosed with stage IVa lung adenocarcinoma on pleural fluid cytology. He received six cycles of carboplatin and pemetrexed, then maintained on pemetrexed. He had disease progression after nine cycles of pemetrexed and was switched to nivolumab. He progressed after five cycles of nivolumab. Core biopsy of the lung mass revealed SCLC. Case 2, a 68-year-old man was diagnosed with two primary NSCLCs and underwent resection. He had recurrence after several months and was treated with four cycles of carboplatin, paclitaxel, and pembrolizumab on clinical trial, with partial response. He was continued on pembrolizumab and had disease progression after 30 cycles. Biopsy of the new lesions showed SCLC. Discussion: Histologic transformation from NSCLC to SCLC can be explained by the presence of a common cell precursor. Proposed molecular mechanisms include loss of RB1, TP53 mutations, and MYC amplification. The distinction between transformation and mixed histology tumors is challenging, especially when pathologic material used for the initial diagnosis is limited. The possibility of a second metachronous primary lung cancer cannot be excluded in our cases. Conclusion: Histologic transformation with PD-1 inhibitors could be under-recognized. Disease progression should prompt re-biopsy to uncover new histology and change in treatment. Future studies are needed to elucidate mechanisms and predictors of transformation.