Multi-omics Study On The Pathogenesis Of Renal Angiomyolipoma Associated With Tuberous Sclerosis Comple

Background:Tuberous sclerosis complex(TSC)is an autosomal dominant genetic disorder caused by mutations in the TSC1 or TSC2 gene that affects multiple organs throughout the body.Rupture and bleeding of tuberous sclerosis complex associated renal angiomyolipoma(TSC-RAML)is the most common cause of death among adult TSC patients.Everolimus is currently the only drug approved for the treatment of surgically unresectable TSCRAML patients.However,around 30%patients still have poor responses to everolimus,which requires deeper exploration of the TSC-RAML pathogenesis.Purposes:Ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS)was applied to profile TSC-RAML plasma proteomics and metabolomics,and TSC-RAML pathogenesis was explored by tissue multiomics techniques.Methods:Plasma samples from TSC-RAML patients before and after everolimus treatment,sporadic angiomyolipoma(S-AML)and renal cyst(CY)patients were collected respectively,from September 2016 to September 2017.UPLCMS was used to detect proteins and metabolites within plasma samples to find biomarkers for TSC-RAML diagnosis and prediction of everolimus efficacy.Differentially expressed proteins and metabolites were enrolled into further pathway enrichment analyses.Tumor and corresponding paratumor normal tissues of TSC-RAML and S-AML,surgically resected from September 2016 to January 2022 were collected.Through multi-omics profiling,including proteomics,metabolomics,bulk and single cell RNA transcriptomics,the dysregulated metabolome and altered tumor microenvironment of TSC-RAML were explored.Results:Finally,plasma samples from 29 pre-treatment and 25 posttreatment TSC-RAML,29 S-AML and 27 CY patients were enrolled.A total of 997 plasma proteins and 517 metabolites were detected with untargeted UPLC-MS.Plasma proteomics analysis identified a total of 34 protein markers,including pre-melanosome protein(PMEL),which could be used for TSC-RAML diagnosis and prognostic prediction,and pathway enrichment analysis showed that TSC-RAML specific plasma proteins were mainly enriched in angiogenesis,smooth muscle proliferation and migration and glycolipid metabolism pathways.Plasma metabolomics identified 13 potentially differential and prognostic metabolic markers including SAdenosylmethionine(SAM).Pathway enrichment analysis showed that arginine and methionine metabolism pathway was up-regulated,while tryptophan metabolism pathway was down-regulated within TSC-RAML plasma.Tissue proteomics suggested that TSC-RAML specific proteins were mainly enriched in small molecule metabolism,amino acid and fatty acid metabolism pathways.Tissue metabolomics showed that purine metabolism,glycolipid metabolism and metabolism of various amino acids were upregulated within TSC-RAML tissues.Tissue RNA transcripts analysis showed that the angiogenesis and oxidative phosphorylation within TSCRAML tumors were upregulated.In addition,multiple cellular component alterations within tumor microenvironment,including increased proportions of fibroblasts and massive infiltration of macrophages,might be involved in the tumorigenesis and TSC-RAML development.Conclusions:TSC-RAML patients have characteristic plasma proteome and metabolome.Plasma proteins and metabolites represented by PMEL and SAM demonstrated roles of differential diagnosis and efficacy prediction.There existed not only abnormalities in metabolism including purine and amino acid metabolism,but also changes in the proportion of fibroblasts and macrophages within TSC-RAML microenvironment.