Vasoactive Intestinal Peptide Receptor Antagonists Potentiate Activation of Human T cells and Enhance Survival Rates in Murine Leukemia Models Open Access
Cohen, Jamie (Spring 2025)
Abstract
Acute Myeloid Leukemia is an aggressive cancer of hematopoietic progenitor cells in the bone marrow that spreads into the blood. Vasoactive intestinal peptide (VIP) is an immunoregulatory peptide that promotes cancer growth in solid tumors, including lung cancer, PDAC, and breast cancer. A subset of AML clinical cases over-express VIP, and the VPAC1 receptor is upregulated in T cells from AML patients. VIP reduces T cell proliferation in vitro, suggesting the VIP-signaling pathway may be an immune checkpoint in AML. Previous experimentation showed that inhibiting the VIP-signaling pathway with a VIP-receptor (VIPhyb) peptide antagonist enhanced the activity of anti-leukemic T cell responses in murine models of acute myeloid leukemia (Petersen et al., 2017). AlphaFold technology identified novel peptide sequences with high predicted binding affinity in silico to human VPAC1 and VPAC2 VIP receptors. We tested ANT308, a peptide antagonist of the VPAC-1 and VPAC-2 receptor with a positively charged N-terminus, and additional peptide sequences, for their ability to promote T cell activation in vitro and potentiate in vivo anti-leukemia activity in mouse models. We demonstrated that ANT308 enhanced the activation of CD4+ and CD8+ T cells and down-regulated expression of the PD-1, another immune checkpoint receptor, that is targeted in treating solid tumor malignancies. Taken together, our data indicate that novel VIP-receptor antagonists can potentiate the activation of human T cells and anti-leukemia activity in mice. These data indicate novel VIP-receptor antagonists are promising candidates for drug development in leukemia and combined with other immune check-point inhibitors for treating solid tumors such as PDAC.
Table of Contents
Table of Contents
Introduction
Background
Treatment History and Developments
Aims of Study
Results
CD3/CD28/CD2 Activator potentiates T cell activation.
Overexposure to CD3/CD28/CD2 Activator Cocktail leads to hyperactivation, exhaustion, and apoptosis of T cells.
Pooling donor T cells post isolation is essential for reproducible T cell activation.
ANT308 treatment potentiates activation of healthy donor T cells.
ANT308 inhibits VIP cell signaling pathway, maintaining T cell activation in vitro.
ANT308 potentiates activation and cytotoxic activity in AML donor T cells.
PEGylation of antagonist peptides enhances T cell activation; Initial Testing of Fc-fusion ANT308 resulted in minimal enhancement of T cell activation.
ANT308 potentiates T cell activation to the highest degree while ANT1201 induces moderate levels of T cell proliferation via CD69 expression.
ANT308 potentiates T cell activation to a limited degree via intracellular protein expression.
Treatment with 100ug of ANT308 enhances survival in B6 leukemic mice, 20ug of ANT308 and ANT1201 enhances T cell activation
Discussion
Summary of Results
Implications
Future Directions
Materials and Methods
In Vitro T cell Isolation, Staining, Flow Cytometry:
Western Blot:
PBMC, Cell lines and Mice:
Mice Leukemia Cell Injection:
ANT308 and ANT1201 Administration:
Conflict of Interest:
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References
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