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Principal Investigator: Wolfgang Beckabir
Organization: UNIV OF NORTH CAROLINA CHAPEL HILL
Fiscal Year: 2024
Award: $53,974
Funding agency: National Cancer Institute
Project Summary/Abstract
Bladder cancer is a prevalent and deadly cancer, with over 80,000 new cases and 17,000 deaths annually in the
United States. Advanced bladder cancer has only a 15% 5-year survival rate. One of the most effective
treatments for advanced bladder cancer is immune checkpoint blockade (ICB), but only 20-30% of patients with
advanced bladder cancer respond and most responses are not enduring. A promising new treatment identified
by our group to improve bladder cancer ICB response is the selective class 1 histone deacetylase inhibitor
entinostat. In a mouse model, entinostat plus anti-PD-1 (αPD-1) ICB induced complete, enduring responses in
67% of mice. Entinostat decreased intratumoral M-MDSC and Treg populations, decreased tumor single-
nucleotide variant (SNV) neoantigen burden in vivo, increased expression of some SNV neoantigens in vitro,
and increased T cell specificity for these neoantigens in vivo. However, much of the mechanism behind response
to entinostat plus αPD-1 is unknown, particularly how entinostat decreases immunosuppressive populations and
affects expression of the neoantigen landscape. Understanding this mechanism is important to predict which
patients will respond and to potentially improve responses through antigen-directed therapy. We hypothesize
that entinostat-induced ICB response is driven by increased M-MDSC differentiation, decreased M-MDSC
migration, and increased expression of suppressed immunogenic neoantigens, augmenting response to
neoantigen vaccination. I will investigate two components driving response to entinostat plus αPD-1: M-MDSCs
and neoantigens. The training in computational and wet lab immunology, tumor biology and genetics, orthotopic
murine tumor models, and translational research with a clinical trial, will assist me in becoming an independently
funded physician-scientist leading a cancer immunology research lab and caring for bladder cancer patients.
In our M-MDSC-focused Aim 1, I will perform flow cytometry and transwell migration assays with M-MDSCs from
orthotopic bladder cancer model tumors to assess whether M-MDSC differentiation and migration are affected
by entinostat treatment. I will conduct immunofluorescence staining of human tumors from the LCCC1827
entinostat window trial (NCT03978624) to assess whether adding entinostat to ICB treatment decreases M-
MDSCs in humans. The Vincent Lab has developed LENS, a software platform to identify neoantigens from
multiple genomic sources. In our neoantigen expression-focused Aim 2, I will use LENS to identify all the
neoantigens in 3 murine bladder cancer lines developed by the Kim Lab, test T cell neoantigen specificity by
high-throughput ELISPOT, and use statistical modeling to predict neoantigens immunogenicity. I will also
validate whether entinostat-induced immunoediting occurs in human tumors from LCCC1827. In our neoantigen
vaccination-focused Aim 3, I will test whether neoantigen vaccination improves tumor response to entinostat
plus αPD-1. I will treat orthotopic tumors, measure their growth, and perform flow cytometry and single cell
RNAseq to measure non-exhausted neoantigen-specific CD8+ T cell abundance.
Terms: <Affect><Antigens><Automobile Driving><Bio-Informatics><Bioinformatics><Bladder Cancer><CD8 Cell><CD8 T cells><CD8 lymphocyte><CD8+ T cell><CD8+ T-Lymphocyte><CD8-Positive Lymphocytes><CD8-Positive T-Lymphocytes><Cancer Model><Cancer Patient><CancerModel><Cancers><Caring><Cell Migration Assay><Cessation of life><Checkpoint inhibitor><Clinical Trials><Collaborations><Computer software><Death><Development><ELISPOT><Epigenetic><Epigenetic Change><Epigenetic Mechanism><Epigenetic Process><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Funding><Future><Generalized Growth><Genetic><Genitourinary><Genitourinary system><Genomics><Growth><HD1><HDAC Agent><HDAC inhibitor><HDAC1><HDAC1 gene><Histone Deacetylase 1><Histone Deacetylase Inhibitor><Histone deacetylase inhibition><Human><Immune checkpoint inhibitor><Immunochemical Immunologic><Immunofluorescence><Immunofluorescence Immunologic><Immunologic><Immunological><Immunologically><Immunologics><Immunology><In Vitro><Knowledge><Malignant Bladder Neoplasm><Malignant Neoplasms><Malignant Tumor><Malignant Tumor of the Bladder><Malignant neoplasm of urinary bladder><Measures><Mice><Mice Mammals><Migration Assay><Modeling><Modern Man><Murine><Mus><Oncology><Oncology Cancer><Patients><Physicians><Population><Position><Positioning Attribute><Probabilistic Models><Probability Models><Prognosis><RNA Seq><RNA sequencing><RNAseq><RPD3-Like 1><RPD3L1><Reduced Potassium Dependency 3, Yeast, Homolog-Like 1><Regulatory T-Lymphocyte><Research><Sampling><Scientist><Single Base Polymorphism><Single Nucleotide Polymorphism><Software><Source><Specificity><Staining method><Stains><Statistical Models><Survival Rate><T Cell Specificity><T cell response><T-Cell Immunologic Specificity><T-Cells><T-Lymphocyte><T8 Cells><T8 Lymphocytes><Testing><Tissue Growth><Training><Translational Research><Translational Science><Treg><Tumor Antigens><Tumor Biology><Tumor Volume><Tumor-Associated Antigen><United States><Urinary Bladder Cancer><Urinary Bladder Malignant Tumor><Urogenital><Urogenital System><Work><aPD-1><aPD1><anti programmed cell death 1><anti-PD-1><anti-PD1><anti-programmed cell death protein 1><antiPD-1><antiPD1><cancer antigens><cancer diagnosis><cancer immunobiology><cancer immunology><cancer microenvironment><career><check point blockade><checkpoint blockade><developmental><driving><effective therapy><effective treatment><enzyme linked immunospot assay><epigenetically><experience><flow cytophotometry><immune check point blockade><immune check point inhibitor><immune checkpoint blockade><immunogen><immunogenic><immunogenicity><improved><in vivo><malignancy><men><migration><mouse model><murine model><neo-antigen><neo-antigen targeted vaccination><neo-antigen vaccination><neo-epitopes><neoantigen targeted vaccination><neoantigen vaccination><neoantigens><neoepitopes><neoplasm immunology><neoplasm/cancer><ontogeny><regulatory T-cells><response><scRNA-seq><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single nucleotide variant><single-cell RNA sequencing><statistical linear mixed models><statistical linear models><subcutaneous><subdermal><thymus derived lymphocyte><transcriptome sequencing><transcriptomic sequencing><transcriptomics><translation research><translational investigation><treatment effect><tumor><tumor immunology><tumor microenvironment><tumor-specific antigen><ultrasound><αPD-1><αPD1>