| Background:TSC is an autosomal dominant disease characterized by the growth of benign tumors in multiple organs and systems.Inactivating mutations of TSC1 or TSC2 gene could be found in most TSC patients.Abnormal activation of mTOR pathway caused by these mutations is the major mechanism of TSC pathogenesis.Therefore,mTOR inhibitors are used as the etiological treatment for TSC-RAML.Approximately 21-30%of TSC-RAML patients have a poor response to the mTOR inhibitor treatment.New therapeutic targets for TSC-RAML are needed.Methods:We acquired 3 TSC-RAML tissue and paired para-tumor kidney tissue from TSC-RAML patients who had surgical indications.Single-cell transcriptome sequencing was performed in the TSC-RAML samples and three para-tumor kidney samples.Patient-derived tumor models were constructed using the conditional programming culture(CRC)technique.A drug panel containing 35 targeted drugs was used to test the drug sensitivity of TSC-RAML CRCs.Results:Smooth muscle cells accounted for 20.38%of all TSC-RAML cells.The proportions of smooth muscle cells and M2 macrophages significantly increased in TSC-RAML compared with TSC kidney samples(P=0.0009 and 0.0011,respectively).In TSC-RAML smooth muscle cells,885 genes were upregulated.These genes were enriched in the myogenesis,response to wounding,response to growth factor,and signaling by receptor tyrosine kinase pathways.Intercellular communication analysis showed that smooth muscle cells were the major transmitters and receivers of FGF signaling in TSC-RAML.In TSC-RAML M2 macrophages,94 genes were upregulated.These genes were enriched in negative regulation of immune system processes,response to wounding and myeloid leukocyte migration pathways.Four CRCs were constructed using the tissue of TSC patients.The pathogenic mutation and STR profile between the CRCs and tissue were identical,and the SNV overlap was 95.8%and 97.0%,respectively.The sensitivity to everolimus of CRCs was similar to the clinical manifestations of TSC-RAML patients.Selective FGFR inhibitor showed a high drug sensitivity score(DSS)in the TSC-RAML CRCs(26.90 on average),and the DSS of FGFR inhibitors were low in the TSC kidney CRCs(2.23 on average).Conclusion:Smooth muscle cells with high ACTA2 and PMEL expression are the key cellular components of TSC-RAML.Smooth muscle cells achieve growth signaling via the FGFR pathway.Targeting FGFR could significantly inhibit the viability of TSC-RAML cells and have a relatively low influence on TSC kidney cells,suggesting it may be a novel therapy for TSC-RAML. |
