Collaborating Investigator: Jeffrey Weber
Affiliation: Deputy Director of the Perlmutter Cancer Center, NYU Langone

Funding source: NIH
GrantNumber: R01CA244936 
Project period: 07/02/20 – 06/30/25 

Significance

Jeff Weber is the deputy director of the NYU Perlmutter Cancer Center and leads a multidisciplinary team of medical and surgical oncologists, dermatologists, and pathologists to treat patients with melanomas ranging from the most common to the most complex. He is the codirector of the Melanoma Research Program and oversees work in experimental therapeutics. His clinical and research interests lie in the field of immunotherapy for cancer, and is at the forefront of new ideas in immunotherapy for treating patients with melanoma. The Weber lab has access to samples from melanoma patients treated with nivolumab and has ongoing participation in immunotherapy clinical trials, most recently the Merck/Moderna neoepitope vaccine trial. CP#3 is a continuation of a productive CP which has led to multiple publications including recent, Clin Cancer Res, 2020, and Nature Communications 2022 (see #44 and #16 in the Publication List; Overall Section). 

Approach

Aim 1: we propose to subject available frozen tumors from twenty melanoma patients from which we have matching peripheral blood mononuclear cells to whole exome sequencing and RNA Seq analysis to define neo-antigens using established software algorithms. In aim 2, we propose to expand optimal PD-1 blockade-inducing neoantigen-specific T cells for modelled on what would be an ACT trial.

As part of this Collaborative Project with TR&D#1, effector cells will then be rapidly expanded after exposure to neo-antigen peptide-pulsed nanoparticle-based aAPC that express anti CD28 and HLA on their surface, using all 20 peptides. The concentration of peptides pulsed onto the aAPC will be varied by a factor of 100 around 1 micromolar to test the peptide concentration that results in the best yield of antigen specific T cells. We will establish the feasibility of simultaneously expanding multiple neo-antigen specific T cells from at least 20 patients with melanoma using available peripheral blood cells taken after PD-1 blockade from a previous clinical trial with nivolumab. The feasibility of this process will be defined at several levels. First, we wish to be able to show that at least 2 of the top 20 predicted binding peptides can be pulsed onto aAPC and generate T cell populations that recognize HLA matched melanoma cell lines. The second level of feasibility involves expansion of the resulting T cells. Feasibility would be defined as expansion of T cells that are predominantly (more than 75%) CD8 positive at least 1000-fold in a 2-week period using peptide pulsed MP-based aAPC obtained through this CP from three quarters of patients tested. Optimal conditions of aAPC concentration and ratio to PBMC, peptide concentration for pulsing, and cytokine concentration will be identified that primarily promote expansion of a central memory- like phenotype of CCR7+, CD45RA- CD8+ T cells. Subsequent conditions will be selected that allow the expansion of cells that retain high effector functions, including proliferation, IFN production and granzyme mobilization.

Push-Pull relationship

The NCBIB will send the Weber lab MP-based class I and class II HLA-based aAPC, including HLA A2 and also class II HLA-based MP with either HLA DR4 and DP4. They will study expansion of shared tumor antigen, such as Mart-1 and GP-100, T cells from patients with melanoma who have received various types of immunotherapy and also neoepitopes-specific T cell responses. Their analysis will include high-dimensional 50-color flow cytometry, as well as modelling expansion of T cells for potential Adoptive Cell Therapy, ACT, trials. Pull: Dependent on their findings, we will modify the aAPC to help optimize formulations and develop expansion SOP to optimize TCM, central memory T cell production, in mock up for ACT-based clinical trials of both CD8 and CD4 T cells for melanoma patients.