Multiplexed relative and absolute quantitative immunopeptidomics reveals MHC I repertoire alterations induced by CDK4/6 inhibition
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Abstract
Peptides bound to class I major histocompatibility complexes (MHC) play a critical role in immune cell recognition and can trigger an antitumor immune response in cancer. Surface MHC levels can be modulated by anticancer agents, altering immunity. However, understanding the peptide repertoire's response to treatment remains challenging and is limited by quantitative mass spectrometry-based strategies lacking normalization controls. We describe an experimental platform that leverages recombinant heavy isotope-coded peptide MHCs (hipMHCs) and multiplex isotope tagging to quantify peptide repertoire alterations using low sample input. HipMHCs improve quantitative accuracy of peptide repertoire changes by normalizing for variation across analyses and enable absolute quantification using internal calibrants to determine copies per cell of MHC antigens, which can inform immunotherapy design. Applying this platform in melanoma cell lines to profile the immunopeptidome response to CDK4/6 inhibition and interferon-γ - known modulators of antigen presentation - uncovers treatment-specific alterations, connecting the intracellular response to extracellular immune presentation.
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The MHC class I peptidome of human melanoma cells treated with palbociclib is enriched for tumor-associated antigens and peptides from E2F target proteins involved in cell cycle blockage.
“the MHC class I peptidome of human melanoma cells exposed to the CDK4/CDK6 inhibitor palbociclib is enriched for tumor-associated antigens (TAAs) and peptides from E2F target proteins that are directly implicated in cell cycle blockage (Stopfer et al., 2020).”
From Immunomodulation by targeted anticancer agents (2021) · cites this paper as Stopfer et al., 2020
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Systems analysis uncovers early temozolomide responses and peptide antigens in glioblastomaPreprint
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Control of antigen presentation on MHC-I by a bacterial secretion systemPreprint
Leddy, O. K., McGinn, R. A., Allsup, B. L., Leone, M., Timms, P., Gutiérrez-González, M., Fahad, A. S., Milligan, R., García-Beltrán, W. F., DeKosky, B. J., et al. (2025) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2025.11.19.689332
Learning the Principles of T Cell Antigen Discernment
Bourassa, F. X. P., Achar, S. R., Altan-Bonnet, G. Y., & François, P. (2026) · Annual Review of Biophysics · doi:10.1146/annurev-biophys-021424-013707
Cui, Y., Phuong, K., Temple, H. M., Wisdom, A. J., Abdelfattah, N. S., Spranger, S., & White, F. M. (2026) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.64898/2026.02.23.707477
Kinase inhibition rewires the HLA-I immunopeptidome in chronic myeloid leukemia
Venafra, V., Wahle, M., Massacci, G., Bica, V., Chiusolo, P., Mougiakakos, D., Böttcher, M., Fischer, T., Perfetto, L., Mann, M., et al. (2026) · iScience · doi:10.1016/j.isci.2026.115944
- Authors
- Stopfer, L. E., Mesfin, J. M., Joughin, B. A., Lauffenburger, D. A., & White, F. M.
- Affiliations
- Koch Institute for Integrative Cancer Research At MIT (US)
- Massachusetts Institute of Technology (US)
- Corresponding author
- White, Forest M.
- Venue
- Nature Communications
- Published
- 2020-06-02
- Volume
- 11
- Issue
- 1
- Pages
- 2760-2760
- ISSN
- 2041-1723 (ISSN-L)
- PMC
- PMC7265461
- OpenAlex
- W3031716951
- Cited by
- 109
- References
- 79
- Citations by year
- 2026: 17 · 2025: 21 · 2024: 11 · 2023: 23 · 2022: 14 · 2021: 16 · 2020: 6 · 2019: 1
- FWCI
- 4.76
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Nature Communications (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- DSpace@MIT (Massachusetts Institute of Technology) (submittedVersion, cc-by-nc)
- Funders
- Melanoma Research Alliance (565436)
- Array BioPharma
- Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology
- National Institutes of Health (CA210180)
- National Heart, Lung, and Blood Institute (T32HL105373)
- National Cancer Institute (U54CA210180, P30CA014051)
- National Institute of Environmental Health Sciences (T32ES007020)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › vaccines and immunoinformatics approaches
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Health Sciences › Medicine › Oncology › Cancer Immunotherapy and Biomarkers
- Keywords
- immunopeptidomics, CDK4/6 inhibition, absolute quantification, melanoma, interferon-gamma, antigen presentation, cancer immunotherapy
- MeSH terms
- Antigens, Cell Line, Humans, Immunotherapy, Interferon-gamma, Peptides, Mass Spectrometry, Histocompatibility Antigens Class I, Antigen Presentation, Proteomics, Cyclin-Dependent Kinase 4, Cyclin-Dependent Kinase 6
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