Endosome-microautophagy targeting chimera (eMIATAC) for targeted proteins degradation and enhance CAR-T cell anti-tumor therapy.

in Theranostics by Kunjian Lei, Jingying Li, Zewei Tu, Chuandong Gong, Junzhe Liu, Min Luo, Wenqian Ai, Lei Wu, Yishuang Li, Zhihong Zhou, Zhihao Chen, Shigang Lv, Minhua Ye, Miaojing Wu, Xiaoyan Long, Xingen Zhu, Kai Huang

TLDR

  • The study developed a new way to get rid of abnormal proteins in the body. They used a system called eMIATAC that works by taking the abnormal protein and putting it in a special bag that will take it to the garbage disposal. The eMIATACs were very good at getting rid of the abnormal protein and could be used to treat cancer and genetic diseases. The study also showed that eMIATAC could help CAR-T cell therapy for tumors by getting rid of proteins that make the CAR-T cells less effective.

Abstract

Since oncogene expression products often exhibit upregulation or abnormally activated activity, developing a technique to regulate abnormal protein levels represent a viable approach for treating tumors and protein abnormality-related diseases.We first screened out eMIATAC components with high targeted degradation efficiency and explored the mechanism by which eMIATAC induced target protein degradation, and verified the degradation efficiency of the target protein by protein imprinting and flow cytometry. Next, we recombined eMIATAC with some controllable elements to verify the regulatable degradation performance of the target protein. Subsequently, we constructed eMIATAC that can express targeted degradation of AKT1 and verified its effect on GBM cell development in vitro and in vivo. Finally, we concatenated eMIATAC with CAR sequences to construct CAR-T cells with low BATF protein levels and verified the changes in their anti-tumor efficacy.we developed a system based on the endosome-microautophagy-lysosome pathway for degrading endogenous proteins: endosome-MicroAutophagy TArgeting Chimera (eMIATAC), dependent on Vps4A instead of lysosomal-associated membrane protein 2A (LAMP2A) to bind to the chaperone Hsc70 and the protein of interest (POI). The complex was then transported to the lysosome by late endosomes, where degradation occurred similarly to microautophagy. The eMIATACs demonstrated accuracy, efficiency, reversibility, and controllability in degrading the target protein EGFP. Moreover, eMIATAC exhibited excellent performance in knocking down POI when targeting endogenous proteins in vivo and in vitro.The eMIATACs could not only directly knock down abnormal proteins for glioma treatment but also enhance the therapeutic effect of CAR-T cell therapy for tumors by knocking down T cell exhaustion-related proteins. The newly developed eMIATAC system holds promise as a novel tool for protein knockdown strategies. By enabling direct control over endogenous protein levels, eMIATAC has the potential to revolutionize treatment for cancer and genetic diseases.

Overview

  • The study aims to develop a system based on the endosome-microautophagy-lysosome pathway for degrading endogenous proteins: endosome-MicroAutophagy TArgeting Chimera (eMIATAC).
  • The study explores the mechanism by which eMIATAC induced target protein degradation and verified the degradation efficiency of the target protein by protein imprinting and flow cytometry. The eMIATACs demonstrated accuracy, efficiency, reversibility, and controllability in degrading the target protein EGFP. Moreover, eMIATAC exhibited excellent performance in knocking down POI when targeting endogenous proteins in vivo and in vitro. The eMIATACs could also enhance the therapeutic effect of CAR-T cell therapy for tumors by knocking down T cell exhaustion-related proteins. The study holds promise as a novel tool for protein knockdown strategies.

Comparative Analysis & Findings

  • The study compared the outcomes observed under different experimental conditions or interventions detailed in the study. The eMIATACs demonstrated accuracy, efficiency, reversibility, and controllability in degrading the target protein EGFP. Moreover, eMIATAC exhibited excellent performance in knocking down POI when targeting endogenous proteins in vivo and in vitro. The eMIATACs could also enhance the therapeutic effect of CAR-T cell therapy for tumors by knocking down T cell exhaustion-related proteins.

Implications and Future Directions

  • The study's findings suggest that the eMIATAC system holds promise as a novel tool for protein knockdown strategies. The system could be further developed to target other endogenous proteins and could be used for the treatment of cancer and genetic diseases. The study also highlights the potential of eMIATAC to enhance the therapeutic effect of CAR-T cell therapy for tumors by knocking down T cell exhaustion-related proteins. Future research could focus on optimizing the eMIATAC system for specific applications and improving its efficiency and specificity.
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