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Predicting response to ICI therapy among patients with renal cell carcinoma (RCC) has been uniquely challenging. We analyzed patient characteristics and clinical correlates from a retrospective single-site cohort of advanced RCC patients receiving anti-PD-1/PD-L1 monotherapy (N = 97), as well as molecular parameters in a subset of patients, including multiplexed immunofluorescence (mIF), whole exome sequencing (WES), T cell receptor (TCR) sequencing, and RNA sequencing (RNA-seq). Clinical factors such as the development of immune-related adverse events (odds ratio (OR) = 2.50, 95% confidence interval (CI) = 1.05–5.91) and immunological prognostic parameters, including a higher percentage of circulating lymphocytes (23.4% vs. 17.4%, p = 0.0015) and a lower percentage of circulating neutrophils (61.8% vs. 68.5%, p = 0.0045), correlated with response. Previously identified gene expression signatures representing pathways of angiogenesis, myeloid inflammation, T effector presence, and clear cell signatures also correlated with response. High PD-L1 expression (>10% cells) as well as low TCR diversity (≤644 clonotypes) were associated with improved progression-free survival (PFS). We corroborate previously published findings and provide preliminary evidence of T cell clonality impacting the outcome of RCC patients. To further biomarker development in RCC, future studies will benefit from integrated analysis of multiple molecular platforms and prospective validation.
Michael F. Murray, MD, FACMG, FACP, Yale University; James P. Evans, MD, PhD, University of North Carolina at Chapel Hill; Misha Angrist, PhD, Duke University; Kee Chan, PhD, University of Illinois at Chicago; Wendy R. Uhlmann, MS, CGC, University of Michigan; Debra Lochner Doyle, MS, LCGC, Washington State Department of Health; Stephanie M. Fullerton, DPhil, University of Washington; Theodore G. Ganiats, MD, University of California at San Diego; Jill Hagenkord, MD, Color Genomics; Sara Imhof, PhD, North Carolina Biotechnology Center; Sun Hee Rim, PhD, MPH, Centers for Disease Control and Prevention; Leonard Ortmann, PhD, Centers for Disease Control and Prevention; Nazneen Aziz, PhD, Kaiser Permanente; W. David Dotson, PhD, Centers for Disease Control and Prevention; Ellen Matloff , MS, MyGene Counsel; Kristen Young, Northwestern University; Kimberly Kaphingst, ScD, University of Utah; Angela Bradbury, MD, University of Pennsylvania; Joan Scott, MS, CGC, Health Resources and Services Administration; Catharine Wang, PhD, Boston University; Ann Zauber, PhD, Memorial Sloan Kettering Cancer Center; Marissa Levine, MD, MPH, University of South Florida; Bruce Korf, MD, PhD, University of Alabama at Birmingham; Debra G. Leonard, MD, PhD, University of Vermont; Catherine Wicklund, MS, Northwestern University; George Isham, MD, HealthPartners; and Muin J. Khoury, MD, PhD, Centers for Disease Control and Prevention
Nitric oxide (NO) produced by endothelial cells in response to cytokines displays anti-inflammatory activity by preventing the adherence, migration and activation of neutrophils. The molecular mechanism by which NO operates at the blood-endothelium interface to exert anti-inflammatory properties is largely unknown. Here we show that on endothelial surfaces, NO is associated with the sulfhydryl-rich protein tissue transglutaminase (TG2), thereby endowing the membrane surfaces with anti-inflammatory properties. We find that tumor necrosis factor-α-stimulated neutrophil adherence is opposed by TG2 molecules that are bound to the endothelial surface. Alkylation of cysteine residues in TG2 or inhibition of endothelial NO synthesis renders the surface-bound TG2 inactive, whereas specific, high affinity binding of S-nitrosylated TG2 (SNO-TG2) to endothelial surfaces restores the anti-inflammatory properties of the endothelium, and reconstitutes the activity of endothelial-derived NO. We also show that SNO-TG2 is present in healthy tissues and that it forms on the membranes of shear-activated endothelial cells. Thus, the anti-inflammatory mechanism that prevents neutrophils from adhering to endothelial cells is identified with TG2 S-nitrosylation at the endothelial cell-blood interface.
