Characterization of microsatellite instability (dMMR/MSI-H) and mutational landscape in a large contemporary cohort of upper tract urothelial cancer (UTUC) patients.

Abstract

465 Background: UTUC is a rare genitourinary malignancy and a number of studies, limited by small sample sizes, have attempted to characterize its mutational landscape. Because immunotherapy is commonly used for this disease type, we evaluated the prevalence of microsatellite instability and characterized the mutational landscapes of UTUC in a large contemporary patient cohort. Methods: UTUC tumor samples were analyzed using next generation sequencing (NGS) (NextSeq, 592 gene panel) or whole exome sequencing (WES) (NovaSeq) (Caris Life Sciences, Phoenix, AZ). Mismatch repair status (deficient [dMMR] or proficient [pMMR]) and microsatellite instability status (MSI-high or stable [MSS]) were detected by immunohistochemistry (IHC), fragment analysis, and NGS. Tumor mutational burden (TMB) was measured by counting all somatic mutations found per tumor (high cutoff ≥ 10 mutations per MB). PD-L1 expression was tested by IHC using PD-L1 antibody clones 22c3 (Agilent; positive cutoff CPS ≥ 10) and SP142 (Ventana; positive cutoff ≥ 5% IC). Pathogenic fusion events were detected using whole transcriptome sequencing (NovaSeq). Statistical significance was determined using the Chi-square test and adjusted for multiple comparison. Results: 538 patients with included – median (range) age 71.5 (30-89) years and 37.5% female/62.5% male. Prevalence of dMMR/MSI-H was 3.9% (21/538) and TMB-high was 22.7% (96/423). Significant molecular differences were not detected in primary vs metastatic disease or in male vs female cases. dMMR/MSI-H tumors had higher frequency of TMB-high compared to MSS tumors (100% vs. 19%, p = 0.00003). dMMR/MSI-H tumors also had a higher frequency than MSS tumors for mutations in genes involved in chromatin remodeling (ASXL 82.4%, CREBBP 60%, SMARCA4 40%, KMT2D 95%, ARIDIA 100%, KMT2A 20%, KMT2C 35.3%, NSD1 20%), DNA-damage repair (FANCG 10%, ATM 45%, ATRX 40%) and other biological pathways (RNF43 10%, PTCH1 21.4%, ERBB3 30%, CDKN2A 25%, TSC2 15%, FLNC 15%, HNF1A 20%, CIC 15%, DNMT3A 17.6%); all adjusted p < 0.05. Pathogenic fusions were detected in 3.8% (17/443) cases, with FGFR3 fusion being the most common, occurring in 2.7% (12/443) cases. PD-L1 positivity was identified in 33.2% (133/400) cases tested by 22c3 antibody and 28.4% (89/313) cases tested by SP142 antibody. No difference was seen in PD-L1 positivity between MSI-H/dMMR vs. MSS tumors. Conclusions: In the largest analysis to date, we found a 3.9% prevalence of dMMR/MSI-high rate in UTUC. All dMMR/MSI-H tumors displayed TMB-high. PD-L1 positivity was comparable between dMMR/MSI-H and MSS tumors. dMMR/MSI-H tumors had a significantly higher rate of mutations in genes involved in chromatin remodeling and DDR biological pathways. These results could inform design of targeted therapy trials in UTUC.

Identification of pathogenic germline variants in a large Chinese lung cancer cohort by clinical sequencing

“…In addition, somatic mutations in ARID1A, CDC73, and SMARCA4 gene were co-occurrent with germline mutations of the MMR pathway (Fig. 4B); previous studies have found that mutations in ARID1A [53], CDC73 [54], and SMARCA4 [55] are enriched in microsatellite instable tumors, which supports our findings and demonstrates these genes may be the targets of MMR deficiency. Germline variants can also affect the distribution of somatic mutation loci.…”

First-line Immune Checkpoint Inhibitor Versus Immune Checkpoint Inhibitor With Chemotherapy for Cisplatin-ineligible Metastatic Urothelial Carcinoma: Evidence From a Real-world, Multicenter Analysis

“…Unlike rare germ line diseases in western countries, the UTUC carcinogenic mechanism had been reported with chronic exposure to aristolochic acid (AA) that induces the TP53 fingerprint mutation. [32][33][34][35][36][37] Recent fundamental research has demonstrated that tumors harboring AA mutational signatures are correlated with a higher tumor mutational burden and more predicted tumor-associated neoantigens, implying the potential efficacy of IO in various types of solid tumors. [38][39][40][41][42] By comprehensive whole genomic sequencing analysis of 90 UTUC patients, Lu et al found that UTUC with AA mutational signature had a significantly higher neoantigen burden, increasing tumor-infiltrating lymphocytes and associated with better cancer-specific survival and metastasis-free survival.…”

“…UTUC is well known for its association with Lynch syndrome and microsatellite instability. Unlike rare germ line diseases in western countries, the UTUC carcinogenic mechanism had been reported with chronic exposure to aristolochic acid (AA) that induces the TP53 fingerprint mutation 32–37. Recent fundamental research has demonstrated that tumors harboring AA mutational signatures are correlated with a higher tumor mutational burden and more predicted tumor-associated neoantigens, implying the potential efficacy of IO in various types of solid tumors 38–42.…”

Efficacy of Immune Checkpoint Inhibitors in Upper Tract Urothelial Carcinomas: Current Knowledge and Future Directions

“…Loss of function in the MMR system, either caused by an inherited mutation or a sporadic event, results in microsatellite instability (MSI) throughout the genome [31]. MSI have been found between 3.9% to 20% in UTUC as compared to <1% in UBC [32,33]. High MSI is correlated with a better prognosis, particularly in patients younger than 71 years old with T2-T3N0M0 tumors [34].…”