bluuebunny/crossref_metadata_2025 huggingface dataset:
Glutamine metabolism and VEGF analysis to elucidate and overcome the mechanism of tyrosine kinase inhibitor resistance in renal cell carcinoma.
Authors: Morozumi Kento , Kawasaki Yoshihide , Sato Tomonori , Maekawa Masamitsu , Takasaki Shinnya , Shimada Shuichi , Sakai Takanari , Yamashita Shinichi , Mano Nariyasu , Ito Akihiro
Published: January 2024

464

Background: Clinical practices have demonstrated improvements in prognosis of patients with advanced renal cell carcinoma (RCC) by combination therapies with tyrosine kinase inhibitors (TKIs) and immune checkpoint inhibitors. However, TKI-resistance is inevitable which hinders further improvement in prognosis. In our previous study, we reported regulating glutamine metabolism could bring about re-sensitivity to Sunitinib (Su) in Su-resistant Renal cell Carcinoma (RCC) cells. However, the mechanism of re-sensitivity and whether it applies to other tyrosine kinase inhibitors (TKIs) has been still unknown. Methods: We established Su and Cabozantinib (Cabo) -resistant cells in 3 RCC cell lines (786-O, Caki-1 and ACHN). We analyzed the activity of glutamine metabolism and VEGF pathway, before and after TKI-resistant、furthermore, conducted antitumor effect in vitro and vivo studies to evaluate re-sensitivity to TKI and relationship between VEGF signaling and regulating glutamine metabolism. Results: In all TKI-resistant cells, the overexpression of glutamine metabolism and VEGF signaling were observed. In vitro and vivo study using Su, the regulating glutaminolysis resulted in 40-74% cell-killing effect in 3 Su-resistant cell lines. Whereas the regulating glutaminolysis also resulted in 35-55% cell-killing effect in 3 Cabo-resistant cell lines. Although the antitumor effect was observed in only 786-O among Su-sensitive cells, it was not observed among Cabo-sensitive cells. Moreover, antitumor effect of regulating glutamine metabolism is more remarkable in vivo. When conducting immunostaining of CD31 to evaluate vascular endothelium, angiogenesis was significantly inhibited by regulating glutamine metabolism.Evaluating VEGF signaling in RT-PCR, VEGFR2 expression and VEGF signaling were downregulated, and PTEN upregulated by suppressing glutamine metabolism. Conclusions: By evaluating glutamate metabolism with VEGF analysis in TKI-resistant RCC cells, we have come to understand a part of the phenomenon of re-sensitivity to TKIs. The inhibition of VEGF signaling and its consequent impact on the tumor microenvironment by regulating glutamine metabolism, are considered to be the mechanisms causing re-sensitivity to TKIs in TKI-resistant cells.We could use TKIs more effectively by regulating glutaminolysis to improve prognosis in advanced RCC patients.
The immune characteristic analysis of BAP1 mutated clear cell renal cell carcinoma.
Authors: Wang Zilin , Liang Jiayu , Zeng Hao
Published: January 2024

466

Background: Commonly mutated genes in clear cell renal cell carcinoma include VHL, PBRM1, and BAP1. Although the detection rate of BAP1 mutations in RCC is not high, from an evolutionary perspective. BAP1 mutated renal cell carcinoma tends to have clinical features of higher malignancy, faster disease progression, and poor efficacy of classic drug treatments. Therefore, this study uses our center and TCGA database to analyze the correlation between transcriptome, immune cell infiltration, immune checkpoint expression and other characteristics and the efficacy of immune checkpoint inhibitor-based treatment in patients with BAP1 mutated renal cell carcinoma. Methods: 1. The correlation between the expression of immune-related factors CD4, CD8 and immune checkpoints PD-L1, LAG3 and BAP1 RNA and the expression difference between wild and mutant groups were analyzed through the TCGA database. 2. Collect BAP1 mutant and wild-type patients retrospectively, and verify the expression of CD4, CD8, PD-L1 and LAG3 by immunohistochemistry. 3. The enrichment analysis of the signaling pathway was carried out after the intersection of TCGA and the differential genes of the center. 4. To analyze the clinical curative effect difference between BAP1 wild-type and mutant advanced RCC patients receiving target immunotherapy. Results: The TCGA data confirmed that there was no difference in the expression of PD-L1 RNA between the two groups (P=0.5); while there was a significant difference in the expression of LAG-3 RNA, which was higher in the BAP1 mutant type than in the wild type (P=0.023); 24 cases of BAP1 wild-type ccRCC specimens available in the gene detection database of our center were selected, and the results of immunohistochemistry suggested that there were differences in the expression of LAG3 (p=0.009) between the BAP1 wild-type and mutant types, while PD There was no significant difference in the expression of -L1 (p=0.157). The transcriptome data enrichment analysis results of TCGA and our center database showed that BAP1 was closely related to cytokine signaling, cAMP signaling pathway and immune response signals in the tumor immune microenvironment. In terms of clinical efficacy, the PFS time of BAP1 wild-type RCC patients receiving ICI combined with TKI therapy was significantly longer than that of BAP1 mutant patients (median follow-up time: 23.93 months; median PFS: 24.63 vs. 11.73 months, HR: 0.304, 95%CI: 0.070-1.324, p=0.025). Conclusions: This study found that BAP1 and cytokines, cAMP pathway and immune inflammation-related pathways were significantly enriched at the transcriptome level. IHC results suggested that LAG3 was more highly expressed in patients with BAP1 mutation. Clinical treatment analysis found that PD-1 inhibitor-based immune combination therapy is not effective for patients with BAP1 mutations. In summary, LAG3 may be a potential therapeutic target in the future for patients with BAP1-mutated RCC.
Therapeutic targeting of metabolic dysfunction by amino acid restriction and alternate day fasting in renal cell carcinoma models.
Authors: Orsi Sabrina , Rupert Chris , Chang Musea , Colligan Sean , Jawadwala Abbas , Delle Fontane Ilaria , De Nigris Filomena , Pili Roberto
Published: January 2024

469

Background: The use of immunotherapies and targeted drugs has significantly improved the clinical outcome of kidney cancer patients. However, the inevitable long-term toxicities and development of drug resistance remain a critical challenge for advanced clear cell and non-clear cell renal cell carcinoma (RCC) patients thus highlighting the need for novel therapeutic strategies. Large-scale metabolomic data have associated metabolic alterations with the pathogenesis and progression of RCC. As diet has been reported to potentially modulate tumor metabolism, dietary interventions represent an innovative strategy to treat metabolically dysregulated tumors. However, the biological basis of the success of dietary interventions is unknown, representing a critical roadblock in the use of dietary restrictions in patients. In this study, we investigated the role of amino acid (AA) restriction and alternate day fasting in metabolic and energy dynamics in preclinical models of clear cell and translocation RCC with the overall goal of expanding the understanding of metabolism’s role in tumor progression and response to therapies. Methods: Seahorse, immunofluorescence, metabolomics, transcriptomics, genetic silencing, and pharmacological inhibition were utilized to assess the mechanisms underlying nutrient utilization and metabolic dysfunction in RCC. Patient-derived organoids and murine xenograft models were used to demonstrate the impact of dietary interventions and targeted therapies Results: Our data suggests that oxidative-phosphorylation is the main source of tumor-derived ATP in a subset of ccRCC cells but in all the tRCC cells assessed. AA restriction was associated with decreased oxidative phosphorylation in RCC models with baseline elevated mitochondrial function. Similarly, RCC models with a more glycolytic phenotype had decreased glycolytic function and reduced tumor burden in response to fasting conditions. The results from combining dietary interventions and targeted therapies are ongoing and will be presented. Conclusions: Our data suggests that oxidative-phosphorylation is the main source of tumor-derived ATP in a subset of ccRCC cells but in all the tRCC cells assessed. AA restriction was associated with decreased oxidative phosphorylation in RCC models with baseline elevated mitochondrial function. Similarly, RCC models with a more glycolytic phenotype had decreased glycolytic function and reduced tumor burden in response to fasting conditions. The results from combining dietary interventions and targeted therapies are ongoing and will be presented.
Spatial analysis of the tumor immune microenvironment in papillary renal cell carcinoma.
Authors: Hayes Mitch , Sabitov Kirill , Soupir Alex C. , Peak Taylor , Nguyen Jonathan , Moran-Segura Carlos , Saeed-Vafa Daryoush , Lopez-Blanco Neale , Ramos Echevarria Paola , Guske Christopher , Balasi Jodi , Dhillon Jasreman , Kim Youngchul , Mule James J , Fridley Brooke , Manley Brandon J.
Published: January 2024

471

Background: Spatial analysis of the tumor immune microenvironment (TIME) has yet to be explored in papillary renal cell carcinoma (pRCC). We utilized multiplex immunofluorescence (mIF) and spatial transcriptomics using spatial molecular imaging (SMI) to evaluate TIME properties in pRCC and contrasted these results with clear cell RCC (ccRCC). Methods: Tumor specimens were obtained from localized RCC tumors. mIF was performed on regions of interest (ROIs) selected from matched compartments from tumor, stroma, and tumor/stromal subsets of the interface. Two antibody panels were used for markers against T cells and B cells/macrophages. Marker abundance and clustering differences between pRCC and ccRCC were evaluated across ROIs. Select markers were also explored across pathologic tumor staging in pRCC. The SMI platform used for validation utilized probes against 959 transcripts. Cells were phenotyped using InSituType using the Kidney Cell Atlas as a reference. Cell clustering was quantified by univariate and bivariate Ripley’s K using the spatialTIME package in R. Results: mIF was performed on 1178 ROIs from 16 pRCC tumors and 70 ccRCC tumors. Compared to ccRCC, pRCC immune cell abundance was statistically lower amongst many T cell types and M2-like macrophages (Figure). M1-like macrophages were the only cell line seen at higher levels in interface compartments only. Increased macrophage clustering was observed in pRCC, including doubly positive M2-like macrophages in interface compartments (p=0.001 and 0.007). Higher abundance of CD8+ and FOXP3+ T cells in pRCC was associated with worse clinical stage, but no trend was seen with marker clustering. Four ROIs from 2 pRCC patients underwent SMI validation. On SMI of the tumor compartment, T cells were clustered with other T cells, B cells, and M1 macrophages. Conclusions: Compared to ccRCC, pRCC has fewer T cells and macrophages but more macrophage clustering. Using spatial transcriptomics, we found significant clustering between T cells, macrophages, and B cells in pRCC.
Initial screening efforts for the OPTIC RCC trial.
Authors: Haake Scott Mattox , Beckermann Katy , Chen Yu-Wei , Reddy Anupama , Mar Nataliya , Ornstein Moshe Chaim , Barata Pedro C. , Zhang Tian , Pal Sumanta Kumar , Rathmell W. Kimryn , Rini Brian I.
Published: January 2024

478

Background: OPTIC RCC (NCT05361720) uses RNA sequencing (seq) of tumor tissue to assign therapy in front-line metastatic clear cell renal cell carcinoma (mccRCC) patients. The operational characteristics of tumor tissue procurement, sequencing, cluster assignment and start of therapy are presented. Methods: Patients diagnosed with mccRCC without prior systemic therapy are eligible for enrollment. RNAseq of primary and/or metastatic tumor tissue is performed by Tempus (Chicago, IL) and data analyzed via an automated cloud-based informatics tool. Results: Twenty-three patients have been screened. Twelve patients (52%) had both primary and metastatic tumors submitted for RNAseq, 4 (17%) had primary only, and 7 (30%) had only metastatic tumor available. Three patients (13%) failed screening because no tumors passed RNAseq quality control (QC). When possible, patients were assigned to clusters based on data from metastatic tumor (15/20, 75%). However, 5/20 (25%) of patients were assigned to clusters based on primary tumor sequencing data because the metastatic tumor was not available or failed QC. Eight patients (40%) were assigned to cluster 1/2, 7 (35%) were assigned to cluster 4/5, and 5 (25%) were assigned to cluster 3/6/7. Primary tumors were more likely to be assigned to cluster 1/2 (12/14, 86%) than metastatic tumors (5/10, 50%) (p<0.01, Fisher’s exact test). Of the 9 pts with both primary and metastatic RNAseq, 5 patients had discordant tumor clusters (primary cluster 1/2 and metastatic cluster 4/5 or 3/6/7); 4 patients with concordant tumor clusters (both cluster 1/2). The mean time from consent to cluster assignment was 32 days (SD +/- 18), though this improved over time. Ten patients have started therapy. Conclusions: Patients with mccRCC can be enrolled on clinical trials that utilize tissue-based RNA sequencing biomarkers. Accrual and analysis correlating cluster assignment with treatment response are ongoing. This trial is funded by the DOD Kidney Cancer Research Program (W81XWH-22-1-1033). Clinical trial information: NCT05361720 .
Safety and efficacy of surgery in renal cell carcinoma with supradiaphragmatic tumor thrombus involving cardiac procedures.
Authors: Nicaise Edouard , Shah Yash , Schmeusser Benjamin , Croll Benjamin , Richter Ellen , Hollon McKenzie , Ogan Kenneth , Daneshmand Mani A. , Maithel Shishir K. , Master Viraj A.
Published: January 2024

484

Background: Invasion of the IVC is a unique feature of RCC. Tumor extension into the supradiaphragmatic IVC carries high morbidity and mortality attributable to surgical complexity. It is unclear whether safety and efficacy have been shown to warrant cytoreductive surgery for metastatic RCC with concomitant thrombus. We present outcomes of cytoreductive and non-cytoreductive radical nephrectomy (RN) with supradiaphragmatic tumor thrombectomy (TT) involving cardiac surgery. Methods: We reviewed our nephrectomy database for patients with RCC and supradiaphragmatic tumor thrombus. RN with TT was performed by a single team from urology, surgical oncology and cardiothoracic surgery. Rank Sum for numerical and chi-square for categorical variables were used to test for differences between cytoreductive and non-cytoreductive RN. Kaplan Meier curves estimated cancer-specific survival (CSS) and overall survival (OS). Results: From 2006-2023, 43 patients underwent RN with supradiaphragmatic TT. The tableindicates notable clinical and surgical features. No patient participated in a related clinical trial perioperatively. No case required circulatory arrest. One patient required temporary dialysis and later passed from multi-organ failure. 5-year CSS was 12% vs. 78% (p=0.007) and OS was 11% vs. 53% (p=0.044) for cytoreductive and non-cytoreductive surgery respectively. Conclusions: RN with supradiaphragmatic TT is safe with durable treatment response in patients with non-mRCC. Although safety of cytoreductive RN is comparable, further investigation into the role of neoadjuvant therapy is recommended. [Table: see text]
Rethinking the definition of stage IV disease in adrenocortical carcinoma: Assessing the impact of clinical lymph node positive disease.
Authors: Gao Helen , Lichtbroun Benjamin , Chua Kevin J. , Srivastava Arnav , Pfail John , Golombos David , Jang Thomas L. , Ghodoussipour Saum
Published: January 2024

5

Background: Stage III (pT3N0M0 or pT1-3N1M0) adrenocortical carcinoma (ACC) encompasses both lymph node positive (LN+) and lymph node negative (LN-) disease. However, in other malignancies, prior data demonstrates that LN+ disease portends outcomes similar to metastatic, stage IV disease. We similarly wanted to examine the impact of clinical LN+ disease on oncologic outcomes in ACC. Methods: Patients with clinical stage III and stage IV (M1) disease were identified using the National Cancer Database (NCDB). Stage III patients were stratified as having either LN+ or LN- disease. Kaplan-Meier curves illustrated overall survival of the three groups – stage III LN-, stage III LN+, and stage IV. Further analysis stratified stage III LN+ and stage IV patients by whether or not patients received treatment (systemic therapy, surgery, or both). Results: A total of 917 patients were included in the analysis – 322 stage III LN-, 67 stage III LN+, and 528 stage IV. 3-year OS for patients with stage IV, stage III LN+, and stage III LN – was 15.6% (95% CI, 12.5%-19.4%), 29.4% (95% CI, 19.8%-43.6%), and 48.6% (95% CI, 43.2%-54.6%), respectively. Within the stage III LN+ group 3-year OS was 33.0% (95% CI, 18.5%-58.8%) for those who received treatment and was 27.4% (95% CI, 16.1% - 46.6%) for those who did not. Within the stage IV group 3-year OS was 29.8% (95% CI, 22.7%-39.3%) for those who received treatment and was 10.2% (95% CI, 7.4% - 14.2%) for those who did not. Conclusions: The current staging paradigm of ACC includes LN+ disease as part of the stage III classification. Herein, we see that clinical stage III LN+ disease had a 3-year OS that was more similar to clinical stage IV disease than clinical stage III LN- disease. While the survival curves appear distinct, due to the rarity of the disease and therefore small sample size there was overlapping confidence intervals. In stage IV patients there was a significant difference in OS at 3 years between those who did and did not receive treatment. This same finding was not found in stage III LN + patients.
A novel radiomics-miRNAs integrated model to predict the histopathology of residual retroperitoneal masses in metastatic nonseminomatous germ cell tumors.
Authors: Li Xiangdong , Ding Renjie , Liu Zhuowei
Published: January 2024

507

Background: Patients with metastatic nonseminomatous germ cell tumours (NSGCTs) are treated with cisplatin-based chemotherapy, followed by postchemotherapy retroperitoneal lymph node dissection (pcRPLND) for patients with residual nodal masses >1 cm. However, 40–50% of patients who undergo pcRPLND are found to have fibrosis/necrotic tissue alone, which may result in overtreatment. Previous prediction models were based on a single method. We hypothesized that a radiomics-based model combined with serum micro-RNAs (miRNAs) including miR371 and miR375 could exhibit better prediction performance in histopathological presence of visible germ cell tumor, teratoma, and fibrosis/necrotic residual retroperitoneal masses. Methods: A retrospective review of 105 patients with 145 residual retroperitoneal masses undergoing pcRPLND for metastatic NSGCTs was performed to develop a prediction model. The dataset was split into separate training cohorts for model construction and validation cohorts for internal validation. On contrast-enhanced axial computed tomography images, 145 corresponding regions-of-interest (ROIs) of residual masses were segmented and 1130 radiomic features per ROI were extracted. After stepwise feature reduction based on reproducibility, variable importance, and correlation analyses, a logistic regression model was established on the selected most important features. Serum miRNA were used to optimize the classifier. The area under receiver operating characteristics curve (AUC), calibration curve, and decision-curve analyses (DCAs) were used to evaluated the prediction model. Results: The trained machine learning model achieved an AUC of 0.772 (95% CI: 0.670-0.875) in the training cohort and 0.764 (95% CI: 0.589-0.938) in the validation cohort. When serum miR371 and miR375 were added to the radiomics signature, the AUC of training cohort was improved to 0.900 (95% CI: 0.826-0.973), and the AUC of validation cohort was improved to 0.939 (95% CI: 0.847-1.000). Nomogram including radiomics score (Radscore) and miRNA was also performed, and the calibration curve analysis and the DCAs confirmed the agreement between prediction and observation and the clinical utility of the radiomics-miRNA prediction model respectively. Conclusions: Radiomics-miRNAs integrated model represents a novel tools for improved prediction of the histopathology of residual retroperitoneal masses in metastatic NSGCTs, aiming at reducing overtreatment. External validation and prospective datasets are needed to further validate this model.
High doses of chemotherapy and autologous hematopoietic stem cell transplantation in patients with relapsed/refractory testicular germ cell tumors: Multicenter real-world experience in Argentina.
Authors: Losco Federico , Angel Martin , Sade Juan Pablo , Vitriu Adriana Verónica , Soulé Tomás , Ayala Luis , Gelo Olivia , Sierra Jeremías , Berro Mariano , Castro Martín , Palmer Silvina , Burgos Ruben , Cruset Soledad , Ferini Gonzalo Ariel , Perez Mariel , Wernicke Priscila , Pelayes Tortosa Sofia , Duarte Patricio , Yantorno Sebastián , Foncuberta María Cecilia
Published: January 2024

508

Background: Germ cell tumors (GCTs) are the most common type of cancer in adolescent and young adult males. Most of them are curable with multimodal treatment: platinum-based chemotherapy (CT) + surgery. Patients who fail first-line treatment usually receive salvage CT with response rates between 30 and 50%. High-dose chemotherapy and autologous hematopoietic stem cell transplantation (HDCT) is an effective treatment in this context. Methods: We conducted a multicenter retrospective analysis of patients ≥ 15 years old with relapsed/refractory GCTs who received HDCT in 13 centers of the Argentine Group of Bone Marrow Transplantation and Cellular Therapy (GATMO-TC) from 2010 to 2022. Results: 114 patients were included, with a median age of 32 years (range: 15-58). Most had non-seminoma histology (66%); The primary tumor site was testis (65%), retroperitoneum (17%), and mediastinum (9%). Forty percent (n:46) had an unfavorable prognosis according to the IGCCCG score. Eighty-two % received ≥ 2 lines of CT before HDCT, 27% being platinum-refractory disease (PRD), and 45% received 2 consecutive courses of HDCT (tandem). All patients received a carboplatin-etoposide-based conditioning regimen. The median engraftment time was 12 days (d) (r8;25) for neutrophils and 15 d (r6;58) for platelets. Bacteremia (25%) and Hepatotoxicity (14.9%) were the most frequent G3-4 adverse events. With a median follow-up of 12.6 months (r1-138), 100 pts were evaluable for survival. 12- and 24-months OS was 67%- 64%; and PFS 56%; - 54% respectively. PRD patients had worse OS and PFS. Eighteen patients remained alive and without relapsing beyond 5 years of follow-up. Conclusions: This real-world multicentric study demonstrates that some patients with refractory/relapsed metastatic GCT are curable by HDCT, being an effective and safe feasible treatment with response rates similar to those described in the literature. The superiority of the tandem approach could not be demonstrated due to the heterogeneity of the sample.[Table: see text]
Primary retroperitoneal germ-cell tumours (pR-GCT): Evaluation of treatment outcomes of an international retrospective collaboration (PRIMERE study).
Authors: Giannatempo Patrizia , Sebastiano Nazzani , Taglialatela Ida , Valentina Bernasconi , Mego Michal , Bimbatti Davide , Secondino Simona , Claps Melanie , Biasoni Davide , Catanzaro Mario , Zimatore Matteo , Torelli Tullio , Stagni Silvia , Macchi Alberto , Tesone Antonio , Pedrazzoli Paolo , Basso Umberto , Procopio Giuseppe , Nicolai Nicola
Published: January 2024

510

Background: pR-GCT corresponds to the clinical case of retroperitoneal ascertained GCT without radiological evidence of primary testicular tumor and accounts for up to 40% of extragonadal GCTs. An occult primary testicular tumor is often missed at diagnosis. Treatment modalities and outcomes have not been specifically addressed, preventing robust recommendations. We performed an international call to assess prognosis of treatment outcomes of these patients. Methods: Clinical, pathological and treatment data pts with pR-GCT between 04/1988 and 01/2022 were retrospectively collected across 4 referral centers. Kaplan Meier methods, univariable and multivariable Cox regression models (MCRMs) were used. Results: Ninety-nine patients (median age 37 yrs - IQR: 29-45) were collected. Ninety-three (94%) had histological diagnosis by biopsy or primary retroperitoneal lymph-node dissection (RPLND): 60 (62.5%) had non-seminomatous pR-GCTs (pR-NSGCTs) and 33 (34.4%) had seminomatous pR-GCTs (pR-SGCTs). IGCCCG prognostic allocation was possible in 91 pts: 34 (35.8%) were good, 23 (24.2%) intermediate and 34 (35.8%) poor risk, respectively. All pts underwent cisplatin-based chemotherapy, usually BEP regimen (94.9%). After chemotherapy, 25 (25.3%) pts underwent orchiectomy: viable tumor was present in 8 (32%), burn-out lesions in 6 (24%) and no lesion in 11 (44%) cases. After a median follow-up of 45 mos (IQR:15-90), the 5-years OS was 58%, being 85% in case of pR-SGCT and 46% in case of pR-NSGCT. According to IGCCCG classification 5-years OS was 83.3% for good, 57% for intermediate and 42% for poor risk patients, respectively. No difference in term of OS (60% vs 56%, p-value 0.81) was observed between patient who had or not radical orchiectomy. At MCRMs only IGCCCG poor risk category (HR 3.2, CI: 1.18-8.84, p-value 0.02) was independent predictors of a worse OS. Conclusions: Eventually, a significant proportion of pts with a pR-GCT had a misclassified primary testicular tumor. 5 yrs-OS according to IGCCCG classification is worse than expected. The role of a surgical exploration of the suspected primary tumor remains controversial and the existence of a real category of p-GCT cannot be excluded.
Pre-orchiectomy serum miRNA-371a-3p and 372-3p levels in men with clinically localized testicular masses.
Authors: Baky Fady , Matulewicz Richard , Knezevic Andrea , Ye Fei , Williams Brandon , Bromberg Maria , Benfante Nicole , Valentino Alisa , Kantor Rachel , So Rachel , Hossain Jahwa , Sheinfeld Joel , Funt Samuel A , Feldman Darren R.
Published: January 2024

519

Background: MicroRNA (miRNA) are short noncoding RNAs involved in regulation of gene expression and have shown promise as novel biomarkers in the diagnosis and management of testicular germ cell tumors (GCT). We report the accuracy of our combined miRNA assay in men with Clinical Stage I (CSI) GCTs. We also sought to explore the relationship between miRNA levels and patient and oncologic variables to better understand how these factors may influence miRNA results. Methods: We retrospectively identified patients at Memorial Sloan Kettering (MSK) with a newly identified testicular mass concerning for GCT who were planned for radical or partial orchiectomy and had pre-orchiectomy MSK miRNA Assay (miRNA-371a-3p and miRNA-372-3p, collectively MMA) drawn. All patients had clinically localized disease based on initial staging imaging and normalized traditional serum tumor markers (STM) post-orchiectomy. Specific miRNA levels were determined using stem-loop RT-qPCR (quantitative reverse transcription PCR), as described previously. Patient and disease factors were abstracted from the medical record. Outcomes of interest were sensitivity, specificity, PPV and NPV of MMA, and area under the receiver operating characteristic curve (AUC) of miRNA-371a-3p and 372-3p levels, calculated relative to detection of viable GCT at orchiectomy. We explored the relationship between pre-orchiectomy miRNA-371a-3p and 372-3p levels and various patient and pathologic factors using Spearman correlation and Wilcoxon rank-sum test. Results: Forty-eight patients were included in this study, 39 of whom had viable GCT at orchiectomy and 9 with no cancer or non-GCTs. Thirty-seven patients with GCT had positive pre-orchiectomy MMA results (sensitivity 94.9%, 95% CI: 88-100), while 1 of 9 patients (Leydig cell tumor) without viable GCT had a positive MMA result (specificity 88.9%, 95% CI: 68.4-100). MMA PPV was 97.4% (95% CI: 92.3-100) and NPV was 80% (95% CI: 55.2-100). AUC of miRNA-371a-3p and miRNA-372-3p was 96% (95% CI: 91.0-100) and 92% (95% CI: 84.2-99.9), respectively. Pre-orchiectomy miRNA-371a-3p and miRNA-372-3p levels were positively correlated with tumor size (Spearman coefficient ρ=0.64, 0.63, respectively; p<0.001), and with serum HCG (ρ=0.55, 0.56; p<0.001) and LDH (ρ=0.46, 0.46; p<0.001), but not AFP (ρ=0.19, 0.17; p=0.20, 0.24). Pre-orchiectomy MMA was not associated with other tested pathologic or patient characteristics including age, BMI, renal function, liver function and andrology labs. Conclusions: miRNA 371a-3p and 372-3p are highly sensitive and specific for presence of viable GCT in patients with clinically localized disease. Tumor size was the only studied pathologic factor or patient characteristic correlated with MMA levels.
Unraveling the testicular seminoma tumor microenvironment by single-cell RNA-sequencing.
Authors: Giannikou Krinio , Lafin John T. , Sfakianos John P. , Horowitz Amir , Hopkins Benjamin , Yu Haocheng , Amatruda James F. , Bagrodia Aditya
Published: January 2024

522

Background: Seminomas is a malignant germ cell tumor (GCT) affecting male adults and is characterized by low somatic mutation burden, histological homogeneity, and high survival rate. It is currently unknown what drives tumorigenesis of this curable testicular cancer. Tumor/immune microenvironment (TME) has a vital role in cancer survival and progression with implications in cell death resistance and immune escape. Herein, we aimed to unravel the transcriptomic TME landscape of seminomas and better understand seminoma pathogenesis. Methods: We performed single cell RNA-sequencing (scRNA-seq; 10X-Illumina platform) on surgically resected pure seminomas from four patients. A total of ~24,000 single cells were analyzed using Seurat/Bioconductor R packages and standard means for data visualization. Key findings were validated by in situ hybridization RNA analysis (RNAscope) on histological sections from these samples. Results: T-cells and Natural-Killer cell populations were mostly predominantly enriched in this seminoma cohort, where tumor cells consisted of a distinct subpopulation with limited presence (1%). Other distinct cell populations were also identified for additional immune cell types e.g., B-cells, macrophages/monocytes, endothelial and smooth muscle cells. ScRNA-seq revealed a unique set of signature marker genes (e.g., POU5F1, KHDC3L, DPPA5, NANOS3) highly expressed in seminoma cells which are associated with pivotal cellular processes e.g., developmental pluripotency and germ cell development. Notably, this gene set was also highly differentially expressed (DE) in The Cancer Genome Atlas (TGCA) seminomas (n=73) compared to TCGA non-seminomas (n=38) (FDR<0.01). Amongst top marker genes, KHDC3L, also higher expressed in seminomas compared to normal testis, was validated by RNA-scope. KHDC3L demonstrated the strongest positive correlation with DPPA5 (Spearman/Pearson; R2=0.8, p<0.0001), top #3 ranked DE gene as well as with other top 20 DE genes identified by scRNA-seq. Remarkably, similar strong correlations among other top DE genes were also found in the testicular TGCA cohort. Top DE genes including master transcription factors (e.g., POU5F1, NANOG, SOX4) are involved in the same protein pathway/networks likely suggesting synergic effect/involvement in the same regulatory circuitry. Gene Set Enrichment (GSEA) analyses demonstrated enrichment in immune signatures and pivotal pathways e.g., Wnt/beta catenin signaling and MYC targets. Conclusions: Our analyses revealed novel marker genes and immune signatures, uniquely expressed in seminomas which consist of a homogeneous cell population across all 4 tumors analyzed, clearly distinct from other cell types identified.
Financial toxicity from PARP inhibitors in castrate-resistant prostate cancer.
Authors: Benjamin David Joseph , Lester-Coll Nataniel Hernan , Rezazadeh Arash
Published: January 2024

54

Background: Approximately 5-10% of men with prostate cancer (PC) are eligible for treatment with PARP inhibitors. As an estimated 120,000 men living in the US have metastatic PC, between 6,000-12,000 men may be eligible for treatment with PARP inhibitors. Recently, several PARP inhibitors have been approved as single or combination therapies in the first-line castrate-resistant (CR) setting and later-line setting including: olaparib, olaparib plus abiraterone (AAP), and talazoparib plus enzalutamide. It is unclear if these newly approved therapies may be cost prohibitive for patients and/or health care systems. Methods: Using average sale price data from UpToDate for PARP inhibitors and novel hormone therapies (NHTs), we calculated the total cost of treatment for each single or combination therapy regimen. We used phase II/III clinical trial data to determine the median duration of treatment for each regimen. Results: The three approved PARP inhibitors have total costs in descending order as follows: AAP plus olaparib ($689,813.77), talazoparib plus enzalutamide ($388,390.46), and olaparib ($202,845.22). In comparison, in the PROfound trial, the cost of AAP alone was $192,140.37 and in the TALAPRO-2 trial, the cost of enzalutamide alone was $ 253,780.80. The difference in estimated cost between olaparib plus AAP versus talazoparib plus enzalutamide per patient is $37,970.29. Conclusions: Despite several new approved therapies in BRCA mutated and HRR mutated mPC, the total costs for PARP inhibitors remain prohibitive for health care institutions. The estimated difference between two newly approved PARP plus NHT regimens could translate in a difference of $455,643,480 in healthcare system costs if all eligible U.S. men received treatment with olaparib plus AAP over talazoparib plus enzalutamide. [Table: see text]
Demographic and prognostic factors in papillary transitional cell carcinoma: An analysis of the National Cancer Database.
Authors: Vinayak Sankalp , Samuel Maya , Han Emily , Silberstein Peter T. , Hsia Beau , Hu Robert
Published: January 2024

541

Background: Transitional cell carcinomas account for the majority of bladder cancers, with the papillary subtype comprising most of these cases. The role of race in bladder cancers has been investigated in the past, but there have been no studies that collectively investigated the role of different demographic factors in survival and mortality in papillary transitional cell carcinoma (PTCC). This study analyzes the effects of demographic factors on overall mortality (OM) in patients with PTCC. Methods: A retrospective cohort analysis was conducted using National Cancer Database data from 2004-2019, including 12,300 patients with histologically confirmed PTCC (ICD-O-3=8130). Sex, race (Black, White, and Other), insurance status, and median household income quartile (based on 2016-2020 American Community Survey data) were examined. Kaplan-Meier curves were used to calculate 2, 5, and 10 year survival (2-5-10 YS). Multivariable Cox regression was used to calculate hazard ratios (HR) to identify independent prognostic factors. Statistical significance was set at α=0.05. Results: Females had higher 2-5-10 YS and decreased OM compared to males (HR=0.85, P<.001). White patient survival was higher at 2 years compared to Black patients, but Black patient survival was higher at 5 and 10 years. There was no significant difference in OM between Black and White patients (HR=1.07, P=0.260). Patients of other races had higher 2-5-10 YS and decreased OM compared to White (HR=0.85, P=0.023) and Black patients (HR=0.80, P=0.011). Patients on Medicaid had lower 2-5-10 YS and increased OM compared to patients on private insurance (HR=1.19, P=0.036). Patients on Medicaid had higher 2-5-10 YS than patients on Medicare, but no significant difference in OM (HR=0.87, P=0.091). Patients on private insurance (HR=0.70, P<.001) and Medicare (HR=0.73, P=0.003) had decreased OM compared to uninsured patients. Patients in the fourth income quartile had decreased OM compared to patients in the first (HR=0.82, P<.001), second (HR=0.87, P<.001), and third quartiles (HR=0.90, P=0.003). Conclusions: Male sex, uninsured status, and lower income were associated with increased OM. Black patients and White patients had increased OM compared to patients of other races. Future research should investigate the reasons for these disparities and how to minimize them. [Table: see text]
Disparities in the incidence of next-generation sequencing (NGS) among patients (pts) with advanced or metastatic prostate cancer (mPC) and urothelial carcinoma (mUC).
Authors: Jo Yeonjung , Tripathi Nishita , Roy Soumyajit , Mathew Thomas Vinay , Galarza Fortuna Gliceida , Hage Chehade Chadi , Gebrael Georges , Narang Arshit , Campbell Patrick , Gupta Sumati , Maughan Benjamin L. , Agarwal Neeraj , Swami Umang
Published: January 2024

542

Background: Based on availability of approved agents targeting tumor somatic mutations, current guidelines recommend NGS for Pts with mPC and mUC. However, large cohort studies in mPC and mUC evaluating disparities in NGS testing have not been reported. Herein, we investigated the effect of social determinant of health (SDOH) on disparities in NGS testing in a large real-world dataset of pts with mPC and mUC. Methods: Pts data were extracted from the nationwide Flatiron Electronic Health Record (EHR)-derived de-identified database. Inclusion: new diagnosis of advanced or metastatic (adv/met) PCa and UCa between March 2015 to December 2022 and receipt of cancer therapy (to include only treatment eligible pts). Disparities in incidence of NGS testing after adv/met diagnosis by SES, R/E, region, and insurance were assessed using Fine and Grey’s (FG) modified stratified Cox proportional hazard (PH) model. FG model considered death as a competing risk. Stratification was done by year of adv/met diagnosis (accounting heterogeneity) and R/E (an effect modifier for SES, region, and insurance), assuming different baseline hazards in each stratum. All analysis done using R v.4.2.3. Results: Analytic cohorts comprised 11,936 pts with mPC and 6,490 pts with mUC. Hazard ratios (HR) and 95% confidence intervals (CI) from FG Cox PH models are reported in the table. For both cancer types, pts were more likely to get NGS testing when they were White, non-Hispanic (compared to Black, non-Hispanic), had higher SES, and had commercial insurance. Conclusions: Our data suggest an association between incidence of NGS testing and several SDOH in mPC and mUC. Further studies are needed to elucidate the reasons for these disparities and ways to mitigate them. [Table: see text]
Development of a novel RNA-based fibroblast growth factor receptor response signature (FGFR-PRS) for use in patients with urothelial cancer (UC).
Authors: Pappan Kirk L. , Mayhew Gregory , Davison James M. , Shibata Yoichiro , Eisner Joel Robert , Beebe Kirk , Milburn Michael Vance
Published: January 2024

548

Background: Interest in FGFR-targeted (FGFRi) therapies for UC or pan-tumor use is growing (ongoing clinical studies include erdafitinib (NCT05316155; NCT04172675; NCT03390504; NCT04083976), LOXO-435 (NCT05614739), and pemigatinib (NCT03914794) following accelerated approval of erdafitinib in locally advanced/metastatic (m) post-chemotherapy UC patients with FGFR2/3 (i.e., DNA mutations and fusions) alterations (ALT). However, ALT status may not fully capture all who may show clinical benefit as responses are observed in both ALT (+) and (-) patients. Improved test strategies are needed to identify patients most likely to respond to FGFRi. FGFR-PRS is an RNA-based signature developed to capture the biology of FGFR-active tumors independent ALT status and is intended to be used as a diagnostic (Dx) to identify a broader patient population likely to respond to FGFRi. Methods: Known oncogenic FGFR3 ALTs (S249C, R248C, FGFR3c-Y373C, FGFR3c-G370C, FGFR3:TACC3 and FGFR3:BAIAP2L1) were used as training labels for nearest centroid classifier development as described in Dabney et al, 2005 ( https://doi.org/10.1093/bioinformatics/bti681 ) using 2/3 of data from the TCGA BLCA dataset, predominantly comprised of muscle-invasive bladder cancer patients and a few mUC patients, for training. The resulting 80-gene signature was applied to the remaining 1/3 of TCGA and a separate BACI mUC dataset (Rose et al, 2021; https://doi.org/10.1038/s41416-021-01488-6) as test sets. In vitro drug sensitivity was evaluated using data from the Genomics of Drug Sensitivity in Cancer database (GDSC; https://www.cancerrxgene.org/ ). Differential gene expression (DGE) results were analyzed using WebGestalt 2019 (https://www.webgestalt.org/). Results: Thirteen and 18% of the TCGA and BACI UC cohorts were ALT (+), whereas 49% and 48% were FGFR-PRS (+) independent of ALT status. About half of the patients in either cohort were ALT (-)/ FGFR-PRS (-) and only 1% were ALT (+)/FGFR-PRS (-). FGFR 1-3 selective drugs had strong inverse correlations between FGFR-PRS score and IC50 across 18 GDSC bladder cancer cell lines. FGFR-PRS (+) samples had enhanced co-expression of FGFR3, FOXA1, and SHH and selected multiple differentiation ontologies. Conclusions: FGFR-PRS (+) captured most ALT (+) tumors and an additional 2X more with similar FGFR pathway activation. FGFR-PRS (+) tumors were associated with gene enrichments for ontologies linked to FGFR3 signaling. The correlation of FGFR-PRS score with in vitro FGFRi activity provided initial utility of the signature, which is undergoing clinical evaluation in the ALAMANCE retrospective study of UC patients treated with FGFRi or other standard-of-care therapies. Analytical validation is ongoing to support FGFR-PRS for use as a clinical trial assay and eventual Dx.
Economic burden of radical cystectomy and trimodal therapy for bladder cancer in the United States: Real-world study.
Authors: Williams Stephen B. , Yapici Halit O. , Singhal Puneet K. , Weimer Ian , Pathan Farah , Lodaya Kunal , Li Haojie
Published: January 2024

555

Background: Bladder cancer (BC) is exponentially deadly once it invades muscle, with the highest per patient treatment costs among cancers. Radical cystectomy (RC) and trimodal therapy (TMT) are recommended treatment options for muscle-invasive BC. This study describes patient/treatment characteristics and short- and long-term costs/healthcare utilization (HCU) associated with RC and TMT. Methods: Patients with BC undergoing RC or TMT were identified using Optum claims data (October 2015-April 2023). Incident cases were captured, and individuals diagnosed with a prior BC, secondary bladder neoplasm, and non-bladder malignancies were excluded. Costs and HCU (e.g., inpatient, outpatient, emergency, pharmacy) were evaluated up to 5 years of follow-up. Results: Median [IQR] ages of the RC (N=839) and TMT (N=484) cohorts were 71 [66-76] and 75 [69-81] years, and NCI comorbidity index was 0.50±0.43 and 0.52±0.45, respectively. Median follow-up durations were 30 [19,47] and 32 [20,48] months for RC and TMT patients, respectively. Median cost and HCU per patient in the RC cohort were $70,671 [$55,878-$106,812] with 26 [16-39] visits in 3 months, $103,579 [$71,12-$158,489] with 56 [39-86] visits in 12 months and increased to $211,671 [$138,597-$346,389] with 163 [119-218] visits in 60 months. For TMT, median cost and HCU per patient were $34,612 [$16,705-$64,263] with 19 [12-33] visits in 3 months, $116,259 [$65,300-$188,622] with 72 [46-94] visits in 12 months, and increased to $274,462 [$186,337-$421,534] with 186 [144-238] visits in 60 months (N=72). Outpatient care was the highest cost contributor for both cohorts across all time periods, except the first 3 months for RC, where inpatient care was predominant. Approximately 25% (N= 211) of patients received neoadjuvant chemotherapy before RC. For patients receiving TMT, median radiotherapy fractions delivered was 36 [18-43], and 8.1% underwent salvage RC. Conclusions: The burden of RC or TMT-treated BC patients remains high, suggesting key targets for cost containment to optimize value-based care. However, our costs are lower compared to previous literature, possibly due to 3rd party payer (i.e., closed networks), treatment advancements, and patient selection. Future research should evaluate the cost and HCU implications of guideline-recommended treatments i.e., neoadjuvant chemotherapy followed by RC vs. TMT with advised chemotherapy type/duration for both. [Table: see text]
Subgroup analyses from READY: Real-world data from an Italian compassionate use program (CUP) of avelumab first-line maintenance (1LM) treatment for locally advanced or metastatic urothelial carcinoma (la/mUC).
Authors: Bracarda Sergio , Antonuzzo Lorenzo , Maruzzo Marco , Santini Daniele , Tambaro Rosa , Buti Sebastiano , Carrozza Francesco , Calabrò Fabio , Di Lorenzo Giuseppe , Fornarini Giuseppe , Iacovelli Roberto , Cullurà Daniela , Messina Carlo , Cerbone Linda , Fazzi Gennaro , Venturini Filippo , Colasanto Raffaele , Necchi Andrea , De Giorgi Ugo
Published: January 2024

558

Background: A multicenter CUP provided early access to avelumab 1LM in Italian patients (pts) with Ia/mUC before reimbursement. Real-world pt characteristics and outcomes with avelumab 1LM from READY were reported previously. Here, we report updated data and subgroup analyses of effectiveness with avelumab 1LM. Methods: This prospective, noninterventional CUP included pts with la/mUC who were progression free after 1L platinum-based chemotherapy (PBC; 4-6 cycles, starting avelumab 1LM 4-10 wk after last PBC dose). Pts were enrolled from Jan 18, 2021 to Mar 7, 2022. Avelumab was provided per physician request and after approval by local ethics committees, per Italian compassionate-use regulations. Pts who had a relapse within 12 mo of prior adjuvant or neoadjuvant systemic therapy, including immune checkpoint inhibitors, were excluded. Results: 464 pts were included (78.45/21.55% male/female; median age, 70.0 y [interquartile range, 63.0-76.0]). At data cutoff (July 30, 2023), median follow-up from start of avelumab 1LM in 411 evaluable pts was 20.24 mo (95% CI, 19.78-20.93); median overall survival (OS) and progression-free survival (PFS) from start of avelumab were 26.22 mo (95% CI, 19.97-not estimable [NE]) and 7.63 mo (95% CI, 5.79-9.24), respectively. In pts aged <60 y (n=53), 60-70 y (n=150), and >70 y (n=208), median OS (95% CI) was not reached (NR; 12.86 mo-NE), NR (24.21 mo-NE), and 24.01 mo (16.94-NE), and median PFS was 5.20 mo (2.83-6.71), 7.70 mo (5.26-10.07), and 8.82 mo (6.05-12.93), respectively. In pts who received 1L cisplatin + gemcitabine (n=183) and 1L carboplatin + gemcitabine (n=219), median OS (95% CI) was NR (16.05 mo-NE) and 25.10 mo (19.97-NE), and median PFS was 6.61 mo (5.30-9.18) and 8.42 mo (6.05-12.73), respectively. The table shows OS and PFS in other subgroups defined by best response to 1L PBC and number of 1L PBC cycles received. Conclusions: Real-world outcomes with avelumab 1LM in this CUP in Italy show clinical benefit across various subgroups. These data are clinically relevant and are consistent with other real-world country-based studies and the phase 3 JAVELIN Bladder 100 trial. Findings further support the use of avelumab 1LM as standard of care in pts with la/mUC who are progression free after PBC. [Table: see text]