Novelty in improvement of CAR T cell-based immunotherapy with the aid of CRISPR system.

INTRODUCTION: Chimeric Antigen Receptor (CAR) T cells have tremendous potentials for cancer treatment; however, various challenges impede their universal use. These restrictions include the poor function of T cells in tumor microenvironments, the shortage of tumor-specific antigens and, finally, the high cost and time-consuming process, as well as the poor scalability of the method. Creative gene-editing tools have addressed each of these limitations and introduced next generation products for cell therapy. The clustered regularly interspaced short palindromic repeats-associated endonuclease 9 (CRISPR/Cas9) system has triggered a revolution in biology fields, as it has a great capacity for genetic manipulation. METHOD: In this review, we considered the latest development of CRISPR/Cas9 methods for the chimeric antigen receptor T cell (CAR T)-based immunotherapy. RESULTS: The ability of the CRISPR/Cas9 system to generate the universal CAR T cells and also potent T cells that are persistent against exhaustion and inhibition was explored. CONCLUSION: We explained CRISPR delivery methods, as well as addressing safety concerns related to the use of the CRISPR/Cas9 system and their potential solutions.

Non-surgical In Vivo Germ Cell-mediated Gene Editing by CRISPR Mutagenic Chain Reaction with the Aid of Magnetic Nanoparticles.

Gene therapy via germline cells leads to a permanent genetic modification. The promise of this method is due to its potential for providing a stable therapeutic effect for all who inherit the gene of interest. If germinal therapy is successfully performed, it can eliminate certain diseases from the family and the population. The feasibility of genetic modification in the human germline raises several controversial and bioethical issues. However, gene transfer via male and female germinal cells has been recently explored in animal models. Previous studies have shown that delivering DNA to the testes followed by electroporation is relatively successful in producing germline-mediated alterations. Since this method includes surgical procedures, non-surgical, safer, and less timeconsuming methods would be ideal. Herein, we discuss a potential approach for nonsurgical in vivo germ cell-mediated gene editing by CRISPR mutagenic chain reaction with the aid of magnetic nanoparticles.

Treatment of insulin-dependent diabetes by hematopoietic stem cell transplantation.

Type 1 diabetes (T1D) is an autoimmune disease resulting from the demolition of ß-cells that are responsible for producing insulin in the pancreas. Treatment with insulin (lifelong applying) and islet transplantation (in rare cases and severe diseases), are standards of care for T1D. Pancreas or islet transplantation have some limitations, such as lack of sufficient donors and longtime immune suppression for preventing allograft rejection. Recent studies demonstrate that autologous hematopoietic stem cells (HSC) can regenerate immune tolerance against auto-antigens. Taking advantage of this feature, autologous HSC transplantation (auto-HSCT) is likely the only treatment for T1D that is associated with lasting and complete remission. None of the other evaluated immunotherapies worldwide had the clinical efficacy of auto-HSCT. Therapy with auto-HSCT is insulin-independent rather than reducing insulin needs or delaying loss of insulin production. This review provided the latest findings in auto-HSCT for treatment of T1D.