An inherited mtDNA mutation remodels inflammatory cytokine responses in macrophages and in vivo

Impaired mitochondrial bioenergetics in macrophages can drive hyperinflammatory cytokine production, but whether this is caused by inherited mtDNA mutations is unknown. Here, we address this important question using a multi-omic approach that integrates super-resolution imaging and metabolic analyses to profile macrophages from a mouse model of mitochondrial disease caused by a heteroplasmic mutation (m.5019A>G) in the mitochondrial tRNA for alanine. These m.5019A>G macrophages exhibit defects in respiratory chain complexes and oxidative phosphorylation (OxPhos) due to decreased intra-mitochondrial translation. To adapt to this metabolic stress, mitochondrial fusion, reductive glutamine metabolism, and aerobic glycolysis are all increased. Upon inflammatory activation, type I interferon (IFN-I) release is enhanced, while the production of pro-inflammatory cytokines and oxylipins are restrained in m.5019A>G macrophages. Finally, an in vivo endotoxemia model using m.5019A>G mi..., Proteomic analysis
Cell pellets were lysed, reduced and alkylated in 50 µl of 6 M Gu-HCl, 200 mM Tris-HCl pH 8.5, 10 mM TCEP, 15 mM chloroacetamide by probe sonication and heating to 95ºC for 5 mins. Protein concentration was measured by a Bradford assay and initially digested with LysC (Wako) with an enzyme to substrate ratio of 1/200 for 4 h at 37 ºC. Subsequently, the samples were diluted 10-fold with water and digested with porcine trypsin (Promega) at 37ºC overnight. Samples were acidified to 1% TFA, cleared by centrifugation (16,000 g at RT) and approximately 20 µg of the sample was desalted using a Stage-tip. Eluted peptides were lyophilized, resuspended in 0.1% TFA/water and the peptide concentration was measured by A280 on a nanodrop instrument (Thermo). The sample was diluted to 2 µg/ 5 µl for subsequent analysis.
The tryptic peptides were analyzed on a Fusion Lumos mass spectrometer connected to an Ultimate Ultra3000 chromatography system (both Thermo Scientific, Germany) inc..., , # An inherited mtDNA mutation remodels inflammatory cytokine responses in macrophages and in vivo

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### DATA & FILE OVERVIEW

File list:

Results_gc_length_corrected_WT_NS__VS__5019_NS.csv; Results_gc_length_corrected_WT_LPS_1h__VS__5019_LPS_1h.csv; WT1_NS_S1_R1_001.fastq.gz; WT2_NS_S2_R1_001.fastq.gz; WT3_NS_S3_R1_001.fastq.gz; WT1_LPS_1h_S7_R1_001.fastq.gz; WT2_LPS_1h_S8_R1_001.fastq.gz; WT3_LPS_1h_S9_R1_001.fastq.gz; 5019A_NS_S4_R1_001.fastq.gz; 5019B_NS_S5_R1_001.fastq.gz; 5019C_NS_S6_R1_001.fastq.gz; 5019A_LPS_1h_S10_R1_001.fastq.gz; 5019B_LPS_1h_S11_R1_001.fastq.gz; 5019C_LPS_1h_S12_R1_001.fastq.gz; DIA_proteomics_report.pg_matrix.tsv.xlsx

Relationship between files, if important: Analysed RNA sequencing datasets (CPM) and the fastq files of raw reads for each sample; DIA proteomics (LFQ intensity)

### DATA-SPECIFIC INFORMATION

**Results_gc_length_corrected_WT_NS__VS__5019_NS.csv**

Total: Conditions:

Number of conditions: 2 WT NS 5019 NS
N...,