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(A) mtDNA heteroplasmy in MirT cells on day 5, determined by TaqMan SNP assays. (B) Time course of mitochondrial heteroplasmy following MirT cells generation (n = 3). (C) Correlation between reduction of endogenous mtDNA copy number and donor mtDNA incorporation efficiency. (D) Schematic overview of the adoptive cell transfer (ACT) experiments using OT-1 T cells in MC38 tumor-bearing mice treated with anti-PD-L1 antibody. Created with BioRender.com. (E) Representative OCR traces of Young, Aged, and MirT cells-treated T cells cultured <t>under</t> <t>IL-7</t> maintenance conditions for 5 days following the MirT cells generation protocol. (F) Quantification of spare respiratory capacity (SRC), proton leak, ATP production (G) OCR/ECAR metabolic profiles derived from the OCR and ECAR analyses shown in panel F. (H) Determination of the number of aged OT-1 T cells required to suppress tumor growth following ACT (n = 3–4). (I) Tumor growth following ACT using young or aged OT-1 T cells in combination with anti-PD-L1 antibody (n = 3–4). (J) Comparison of antitumor activity between young OT-1 T cells, aged OT-1 T cells, and MirT cells following ACT in MC38 tumor-bearing mice treated with anti-PD-L1 antibody. Data are presented as mean ± SD in (E) and mean ± SEM in (H–K). P values were determined using one-way ANOVA followed by Tukey’s multiple-comparison test (F) or two-way mixed-design ANOVA followed by Tukey’s multiple-comparison test (J).
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(A) mtDNA heteroplasmy in MirT cells on day 5, determined by TaqMan SNP assays. (B) Time course of mitochondrial heteroplasmy following MirT cells generation (n = 3). (C) Correlation between reduction of endogenous mtDNA copy number and donor mtDNA incorporation efficiency. (D) Schematic overview of the adoptive cell transfer (ACT) experiments using OT-1 T cells in MC38 tumor-bearing mice treated with anti-PD-L1 antibody. Created with BioRender.com. (E) Representative OCR traces of Young, Aged, and MirT cells-treated T cells cultured <t>under</t> <t>IL-7</t> maintenance conditions for 5 days following the MirT cells generation protocol. (F) Quantification of spare respiratory capacity (SRC), proton leak, ATP production (G) OCR/ECAR metabolic profiles derived from the OCR and ECAR analyses shown in panel F. (H) Determination of the number of aged OT-1 T cells required to suppress tumor growth following ACT (n = 3–4). (I) Tumor growth following ACT using young or aged OT-1 T cells in combination with anti-PD-L1 antibody (n = 3–4). (J) Comparison of antitumor activity between young OT-1 T cells, aged OT-1 T cells, and MirT cells following ACT in MC38 tumor-bearing mice treated with anti-PD-L1 antibody. Data are presented as mean ± SD in (E) and mean ± SEM in (H–K). P values were determined using one-way ANOVA followed by Tukey’s multiple-comparison test (F) or two-way mixed-design ANOVA followed by Tukey’s multiple-comparison test (J).
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(A) mtDNA heteroplasmy in MirT cells on day 5, determined by TaqMan SNP assays. (B) Time course of mitochondrial heteroplasmy following MirT cells generation (n = 3). (C) Correlation between reduction of endogenous mtDNA copy number and donor mtDNA incorporation efficiency. (D) Schematic overview of the adoptive cell transfer (ACT) experiments using OT-1 T cells in MC38 tumor-bearing mice treated with anti-PD-L1 antibody. Created with BioRender.com. (E) Representative OCR traces of Young, Aged, and MirT cells-treated T cells cultured <t>under</t> <t>IL-7</t> maintenance conditions for 5 days following the MirT cells generation protocol. (F) Quantification of spare respiratory capacity (SRC), proton leak, ATP production (G) OCR/ECAR metabolic profiles derived from the OCR and ECAR analyses shown in panel F. (H) Determination of the number of aged OT-1 T cells required to suppress tumor growth following ACT (n = 3–4). (I) Tumor growth following ACT using young or aged OT-1 T cells in combination with anti-PD-L1 antibody (n = 3–4). (J) Comparison of antitumor activity between young OT-1 T cells, aged OT-1 T cells, and MirT cells following ACT in MC38 tumor-bearing mice treated with anti-PD-L1 antibody. Data are presented as mean ± SD in (E) and mean ± SEM in (H–K). P values were determined using one-way ANOVA followed by Tukey’s multiple-comparison test (F) or two-way mixed-design ANOVA followed by Tukey’s multiple-comparison test (J).
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(A) mtDNA heteroplasmy in MirT cells on day 5, determined by TaqMan SNP assays. (B) Time course of mitochondrial heteroplasmy following MirT cells generation (n = 3). (C) Correlation between reduction of endogenous mtDNA copy number and donor mtDNA incorporation efficiency. (D) Schematic overview of the adoptive cell transfer (ACT) experiments using OT-1 T cells in MC38 tumor-bearing mice treated with anti-PD-L1 antibody. Created with BioRender.com. (E) Representative OCR traces of Young, Aged, and MirT cells-treated T cells cultured <t>under</t> <t>IL-7</t> maintenance conditions for 5 days following the MirT cells generation protocol. (F) Quantification of spare respiratory capacity (SRC), proton leak, ATP production (G) OCR/ECAR metabolic profiles derived from the OCR and ECAR analyses shown in panel F. (H) Determination of the number of aged OT-1 T cells required to suppress tumor growth following ACT (n = 3–4). (I) Tumor growth following ACT using young or aged OT-1 T cells in combination with anti-PD-L1 antibody (n = 3–4). (J) Comparison of antitumor activity between young OT-1 T cells, aged OT-1 T cells, and MirT cells following ACT in MC38 tumor-bearing mice treated with anti-PD-L1 antibody. Data are presented as mean ± SD in (E) and mean ± SEM in (H–K). P values were determined using one-way ANOVA followed by Tukey’s multiple-comparison test (F) or two-way mixed-design ANOVA followed by Tukey’s multiple-comparison test (J).
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(A) mtDNA heteroplasmy in MirT cells on day 5, determined by TaqMan SNP assays. (B) Time course of mitochondrial heteroplasmy following MirT cells generation (n = 3). (C) Correlation between reduction of endogenous mtDNA copy number and donor mtDNA incorporation efficiency. (D) Schematic overview of the adoptive cell transfer (ACT) experiments using OT-1 T cells in MC38 tumor-bearing mice treated with anti-PD-L1 antibody. Created with BioRender.com. (E) Representative OCR traces of Young, Aged, and MirT cells-treated T cells cultured <t>under</t> <t>IL-7</t> maintenance conditions for 5 days following the MirT cells generation protocol. (F) Quantification of spare respiratory capacity (SRC), proton leak, ATP production (G) OCR/ECAR metabolic profiles derived from the OCR and ECAR analyses shown in panel F. (H) Determination of the number of aged OT-1 T cells required to suppress tumor growth following ACT (n = 3–4). (I) Tumor growth following ACT using young or aged OT-1 T cells in combination with anti-PD-L1 antibody (n = 3–4). (J) Comparison of antitumor activity between young OT-1 T cells, aged OT-1 T cells, and MirT cells following ACT in MC38 tumor-bearing mice treated with anti-PD-L1 antibody. Data are presented as mean ± SD in (E) and mean ± SEM in (H–K). P values were determined using one-way ANOVA followed by Tukey’s multiple-comparison test (F) or two-way mixed-design ANOVA followed by Tukey’s multiple-comparison test (J).
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(A) mtDNA heteroplasmy in MirT cells on day 5, determined by TaqMan SNP assays. (B) Time course of mitochondrial heteroplasmy following MirT cells generation (n = 3). (C) Correlation between reduction of endogenous mtDNA copy number and donor mtDNA incorporation efficiency. (D) Schematic overview of the adoptive cell transfer (ACT) experiments using OT-1 T cells in MC38 tumor-bearing mice treated with anti-PD-L1 antibody. Created with BioRender.com. (E) Representative OCR traces of Young, Aged, and MirT cells-treated T cells cultured under IL-7 maintenance conditions for 5 days following the MirT cells generation protocol. (F) Quantification of spare respiratory capacity (SRC), proton leak, ATP production (G) OCR/ECAR metabolic profiles derived from the OCR and ECAR analyses shown in panel F. (H) Determination of the number of aged OT-1 T cells required to suppress tumor growth following ACT (n = 3–4). (I) Tumor growth following ACT using young or aged OT-1 T cells in combination with anti-PD-L1 antibody (n = 3–4). (J) Comparison of antitumor activity between young OT-1 T cells, aged OT-1 T cells, and MirT cells following ACT in MC38 tumor-bearing mice treated with anti-PD-L1 antibody. Data are presented as mean ± SD in (E) and mean ± SEM in (H–K). P values were determined using one-way ANOVA followed by Tukey’s multiple-comparison test (F) or two-way mixed-design ANOVA followed by Tukey’s multiple-comparison test (J).

Journal: bioRxiv

Article Title: Bidirectional modulation of aging-associated cellular phenotypes by mitochondrial genome replacement

doi: 10.64898/2026.06.26.734763

Figure Lengend Snippet: (A) mtDNA heteroplasmy in MirT cells on day 5, determined by TaqMan SNP assays. (B) Time course of mitochondrial heteroplasmy following MirT cells generation (n = 3). (C) Correlation between reduction of endogenous mtDNA copy number and donor mtDNA incorporation efficiency. (D) Schematic overview of the adoptive cell transfer (ACT) experiments using OT-1 T cells in MC38 tumor-bearing mice treated with anti-PD-L1 antibody. Created with BioRender.com. (E) Representative OCR traces of Young, Aged, and MirT cells-treated T cells cultured under IL-7 maintenance conditions for 5 days following the MirT cells generation protocol. (F) Quantification of spare respiratory capacity (SRC), proton leak, ATP production (G) OCR/ECAR metabolic profiles derived from the OCR and ECAR analyses shown in panel F. (H) Determination of the number of aged OT-1 T cells required to suppress tumor growth following ACT (n = 3–4). (I) Tumor growth following ACT using young or aged OT-1 T cells in combination with anti-PD-L1 antibody (n = 3–4). (J) Comparison of antitumor activity between young OT-1 T cells, aged OT-1 T cells, and MirT cells following ACT in MC38 tumor-bearing mice treated with anti-PD-L1 antibody. Data are presented as mean ± SD in (E) and mean ± SEM in (H–K). P values were determined using one-way ANOVA followed by Tukey’s multiple-comparison test (F) or two-way mixed-design ANOVA followed by Tukey’s multiple-comparison test (J).

Article Snippet: For LNP transfection, T cells were pre-activated for 1 day in TexMACS medium supplemented with recombinant human IL-7 (20 ng/mL; Miltenyi Biotec, 130-095-362) and CD3/CD28 DynabeadsTM (Gibco, 11453D).

Techniques: Cell Culture, Derivative Assay, Comparison, Activity Assay