シャペロンを介したオートファジーは、網膜前駆細胞の恒常性の調節に重要な役割を果たします

PURPOSE: To explore the function and regulatory mechanism of IFITM3 in mouse neural retinal progenitor cells (mNRPCs), which was found to be very important not only in the development of the retina in embryos but also in NRPCs after birth. 方法：公開されたシングルセルシーケンスデータを使用して、MNRPCでのIFITM3式を分析しました。RNA干渉を使用して、IFITM3の発現をノックダウンしました。CCK-8アッセイを使用して、細胞生存率を分析しました。RNA-seq was used to assess mRNA expression, as confirmed by real-time quantitative PCR, and immunofluorescence assays and western blots were used to validate the levels of relative proteins, and autophagy flux assay.リソソームトラッカーは、オルガネラの変化を追跡するために使用されました。 RESULTS: The results of single-cell sequencing data showed that IFITM3 is highly expressed in the embryo, and after birth, RNA-seq showed high IFITM3 expression in mNRPCs.IFITM3がノックダウンされた後、増殖と細胞生存率が大幅に低下しました。細胞膜系とリソソームが劇的に変化し、リソソームが活性化され、ランプ処理された細胞で明らかに凝集しました。LAMP1の発現は、ラパマイシン（RAMP）で治療した後、リソソーム凝集とともに有意に増加しました。さらなる検出により、SQSTM1/P62、HSC70、およびLAMP-2Aがアップレギュレートされたが、LC3A/B発現に有意差は観察されなかった。オートファジーフラックスは生成されませんでした。 結論：IFITM3は、主にシャペロンを介したオートファジー（CMA）を介してMNRPCの生存率と増殖を調節しますが、マクロオートファジー（MA）ではありません。IFITM3 plays a significant role in maintaining the homeostasis of progenitor cell self-renewal by sustaining low-level activation of CMA to eliminate deleterious factors in cells.

PURPOSE: To explore the function and regulatory mechanism of IFITM3 in mouse neural retinal progenitor cells (mNRPCs), which was found to be very important not only in the development of the retina in embryos but also in NRPCs after birth. METHODS: Published single-cell sequencing data were used to analyze IFITM3 expression in mNRPCs. RNA interference was used to knock down the expression of IFITM3. CCK-8 assays were used to analyze cell viability. RNA-seq was used to assess mRNA expression, as confirmed by real-time quantitative PCR, and immunofluorescence assays and western blots were used to validate the levels of relative proteins, and autophagy flux assay. Lysosomal trackers were used to track the organelle changes. RESULTS: The results of single-cell sequencing data showed that IFITM3 is highly expressed in the embryo, and after birth, RNA-seq showed high IFITM3 expression in mNRPCs. Proliferation and cell viability were greatly reduced after IFITM3 was knocked down. The cell membrane system and lysosomes were dramatically changed, and lysosomes were activated and evidently agglomerated in RAMP-treated cells. The expression of LAMP1 was significantly increased with lysosome agglomeration after treatment with rapamycin (RAMP). Further detection showed that SQSTM1/P62, HSC70 and LAMP-2A were upregulated, while no significant difference in LC3A/B expression was observed; no autophagic flux was generated. CONCLUSION: IFITM3 regulates mNRPC viability and proliferation mainly through chaperone-mediated autophagy (CMA) but not macroautophagy (MA). IFITM3 plays a significant role in maintaining the homeostasis of progenitor cell self-renewal by sustaining low-level activation of CMA to eliminate deleterious factors in cells.