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Updated: Jul 28, 2026
Immunodetection of Outer Membrane Proteins by Flow Cytometry of Isolated Mitochondria
Published on: September 18, 2014
Isolation of mitochondrial mutation-specific T cell receptors
Catherine Kirkpatrick1,2, Shanshan Hao1,2, Charles M Quick1
1Department of Pathology, College of Medicine, University of Arkansas for Medical Sciences (UAMS), Little Rock, AR, United States.
Abstract:
Neoantigen-specific T cells play a major role in cancer immunotherapy. Mutated proteins derived from non-synonymous mutations in tumor genomic DNA are the major source of neoantigens. It is conceivable that non-synonymous mutations found in tumor mitochondrial DNA (mtDNA) can also generate neoantigens that can be recognized by T cell receptors (TCRs). However, no tumor mitochondrial mutation-specific TCRs have been reported. As a proof-of-concept study, we obtained 3,798 non-synonymous single-nucleotide mutations across 38 tumor types through The Cancer Mitochondria Atlas project. These mutations were subjected to an epitope prediction algorithm to predict binding affinities for HLA-A*0201. The top 300 candidate peptides were synthesized and screened against 10 healthy donors with homozygous HLA-A*0201 alleles. Mutation-reactive T cells were subjected to single-cell sequencing to identify TCR sequences, followed by validations. Here, four MT-ND2 (A103T) mutation-specific TCRs were isolated from a donor. These TCRs interacted with HLA-A*0201-restricted IMMAMTMKL peptide. Additionally, through a single-cell sequencing approach, we demonstrated that a non-synonymous mutation, MT-CO1 (V274I), could be detected in nearly all tumor cells in a colorectal tumor specimen, whereas other mutations in 2 out of 4 tumors were detected in subclonal populations. This study suggests that isolating tumor mitochondrial mutation-specific TCRs is possible, but some biological barriers need to be considered.
Insights
Researchers identified T cell receptors (TCRs) targeting neoantigens from tumor mitochondrial DNA mutations. This study demonstrates the feasibility of isolating mitochondrial mutation-specific TCRs for cancer immunotherapy, despite potential biological barriers.
Area of Science:
- Oncology
- Immunology
- Genetics
Background:
- Neoantigens derived from tumor genomic DNA mutations are crucial for cancer immunotherapy via T cells.
- Tumor mitochondrial DNA (mtDNA) mutations may also generate neoantigens, but specific T cell receptors (TCRs) targeting them remain uncharacterized.
Purpose of the Study:
- To investigate the potential of non-synonymous mutations in tumor mtDNA as a source of neoantigens.
- To isolate and characterize T cell receptors (TCRs) specific for tumor mitochondrial mutations.
Main Methods:
- Utilized The Cancer Mitochondria Atlas project data to identify 3,798 non-synonymous mtDNA mutations across 38 tumor types.
- Employed epitope prediction algorithms and synthesized candidate peptides for screening against HLA-A*0201 alleles in healthy donors.
- Applied single-cell sequencing to identify and validate mutation-specific TCR sequences.
Main Results:
- Successfully isolated four MT-ND2 (A103T) mutation-specific TCRs that recognized the HLA-A*0201-restricted IMMAMTMKL peptide.
- Detected a non-synonymous MT-CO1 (V274I) mutation in nearly all tumor cells of a colorectal specimen using single-cell sequencing.
- Observed that other mtDNA mutations were present in subclonal populations in 2 out of 4 analyzed tumors.
Conclusions:
- This proof-of-concept study demonstrates the possibility of isolating tumor mitochondrial mutation-specific TCRs.
- Further research is needed to address biological barriers that may influence the efficacy of targeting mtDNA-derived neoantigens in cancer immunotherapy.
