Terminally Exhausted CD8+ Tumor-Infiltrating Lymphocytes and M2-Like Macrophages as Biomarkers of Resistance to First-Line Anti-PD-1 Therapy in Metastatic RCC
Abstract
Background: We propose a whole-slide multiplex immunofluorescence characterization of T-cell exhaustion and of hypothesized modifying factors in the microenvironment of kidney cancer. Roughly 70% of patients presenting a metastatic clear cell renal cell carcinoma (mccRCC) treated with anti-PD-1 do not benefit from therapy due to upfront or secondary resistance. Prior studies unveiled the fact that a high density of non-exhausted T cells (CD8+PD-1+TIM3-LAG3-) was predictive of response to anti-PD-1 therapy in previously treated patients with mccRCC. Despite this, most of the biology underlying immune checkpoint blockade remains unknown, and no biomarker is available in clinical practice to guide the prescription of these therapies. We will study the issue from a new angle with these goals: to better understand the biology, to provide markers of resistance to treatment, and to identify potential new therapeutic targets. Hypotheses: We hypothesize that (1) a high ratio of terminally exhausted CD8+PD1+ tumor-infiltrating lymphocytes (TILs) is predictive of a resistance to anti-PD1 therapy in patients with mccRCC treated with first-line anti-PD1; (2) that CD8+PD-1+ TILs in close proximity to M2-like macrophages are more likely to express terminal exhaustion markers and that a high density of M2-macrophages in close proximity to exhausted T-cell is associated with poorer outcome; and (3) that specific genomic and transcriptomic features are associated with intratumor T-cell exhaustion. Specific Aims: Aim 1. To demonstrate that high levels of intratumoral terminally exhausted T cells are associated with innate resistance to first-line nivolumab treatment in ccRCC. Aim 2: To study the spatial interaction between intratumoral terminally exhausted T cells and tumor-promoting M2-like macrophages in ccRCC. Aim 3: To explore genomic and transcriptomic tumor features associated with high levels of intratumoral terminally exhausted T-cells in ccRCC. Study Design: Using tumor samples from 136 patients enrolled in the phase 2 clinical HCRN, we will perform, on whole-slide images, a multiplex immunofluorescence panel, combined with state-of-the-art image spatial analysis algorithms allowing the analysis of the spatial relationship between each cell subtype. This panel combines markers of CD8 exhaustion (CD8, PD1, TIM3) and M2-like macrophages (CD163). Multispectral images will be scanned using the Vectra Quantitative Workstation (Perkin Elmer) and spectral deconvolution will be performed on Inform (Perkin Elmer). Cell phenotyping and spatial analysis will be performed using Halo (Indica Labs). The generated data will be correlated with Overall Survival, Overall Response Rate, and Progression-Free Survival. Moreover, tumor samples of 106 patients will be extracted for both DNA and RNA, alongside germline DNA from blood, to perform Whole Exome Sequencing and RNA sequencing. PERSONAL STATEMENT Career Goals/Skills Development: My goal is to build a career as a physician-scientist with an independent laboratory focused on the exploration of the microenvironment of kidney cancers and the identification of biomarkers of response and resistance to targeted therapies. I plan on building my skills with hands-on experience optimizing multiplex panels and analyzing the images, as well as interpreting Whole Exome Sequencing and RNA sequencing data, in close contact with my mentor, mentoring younger postdoctoral fellows and research assistants, and attending workshops and online classes. Mentor: Dr. Signoretti is a recognized researcher in the field of translational research in kidney cancer. In addition to being a professor of Pathology, she is a co-leader of the DF/HCC Kidney Cancer Program. Dr. Signoretti has lengthy experience in the exploration of the immune microenvironment of kidney cancer, and her team discovered the first and, so far, only, tissue-based biomarker of response to anti-PD-1 therapy in mccRCC.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Dec 05, 2021
- Source ID
- W81XWH2110808
Entities
People
- Thomas Denize
Organizations
- United States Army