TY - JOUR T1 - Diosgenin enhances liposome-enabled nucleic acid delivery and CRISPR/Cas9-mediated gene editing by modulating endocytic pathways JF - Frontiers in Bioengineering and Biotechnology Y1 - 2023 A1 - Brijesh Lohchania A1 - Abisha Crystal Christopher A1 - Porkizhi Arjunan A1 - Gokulnath Mahalingam A1 - Durga Kathirvelu A1 - Aishwarya Prasannan A1 - Vigneshwaran Venkatesan A1 - Pankaj Taneja A1 - Mohan Kumar KM A1 - Saravanabhavan Thangavel A1 - Srujan Marepally VL - 10 UR - https://doi.org/10.3389%2Ffbioe.2022.1031049 ER - TY - JOUR T1 - The CCR5 Gene Edited CD34+CD90+ Hematopoietic Stem Cell Population Serves as an Optimal Graft Source for HIV Gene Therapy JF - Front. Immunol Y1 - 2022 A1 - Karthik V. Karuppusamy A1 - John Paul Demosthenes A1 - Vigneshwaran Venkatesan A1 - Abisha Crystal Christopher A1 - Prathibha Babu A1 - Manojkumar K. Azhagiri A1 - Annlin Jacob A1 - Veena Vadhini Ramalingam A1 - Sumathi Rangaraj A1 - Mohankumar Kumarasamypet Murugesan A1 - Srujan Marepally A1 - George Varghese A1 - Alok Srivastava A1 - Rajesh Kannangai A1 - Saravanabhavan Thangavel AB -

Transplantation of allogenic hematopoietic stem and progenitor cells (HSPCs) with C-C chemokine receptor type 5 (CCR5) Δ32 genotype generates HIV-1 resistant immune cells. CCR5 gene edited autologous HSPCs can be a potential alternative to hematopoietic stem cell transplantation (HSCT) from HLA-matched CCR5 null donor. However, the clinical application of gene edited autologous HSPCs is critically limited by the quality of the graft, as HIV also infects the HSPCs. In this study, by using mobilized HSPCs from healthy donors, we show that the CD34+CD90+ hematopoietic stem cells (HSCs) express 7-fold lower CD4/CCR5 HIV receptors, higher levels of SAMHD1 anti-viral restriction factor, and possess lower susceptibility to HIV infection than the CD34+CD90- hematopoietic progenitor cells. Further, the treatment with small molecule cocktail of Resveratrol, UM729 and SR1(RUS) improved the in vivo engraftment potential of CD34+CD90+ HSCs. To demonstrate that CD34+CD90+ HSC population as an ideal graft for HIV gene therapy, we sort purified CD34+CD90+ HSCs, treated with RUS and then gene edited the CCR5 with single sgRNA. On transplantation, 100,000 CD34+CD90+ HSCs were sufficient for long-term repopulation of the entire bone marrow of NBSGW mice. Importantly, the gene editing efficiency of ~90% in the infused product was maintained in vivo, facilitating the generation of CCR5 null immune cells, resistant to HIV infection. Altogether, CCR5 gene editing of CD34+CD90+ HSCs provide an ideal gene manipulation strategy for autologous HSCT based gene therapy for HIV infection.

ER - TY - JOUR T1 - Exploring membrane permeability of Tomatidine to enhance lipid mediated nucleic acid transfections JF - Biochimica et Biophysica Acta (BBA) - Biomembranes Y1 - 2018 A1 - Vignesh K. Rangasami A1 - Brijesh Lohchania A1 - Chandrashekhar Voshavar A1 - Harikrishna R. Rachamalla A1 - Rajkumar Banerjee A1 - Ashish Dhayani A1 - Saravanabhavan Thangavel A1 - Praveen K. Vemula A1 - Srujan Marepally KW - Endocytosis AB -

Abstract Intracellular delivery of nucleic acids is one of the critical steps in the transfections. Prior findings demonstrated various strategies including membrane fusion, endosomal escape for the efficient cytoplasmic delivery. In our continuing efforts to improve the nucleic acids transfections, we harnessed cell permeable properties of Tomatidine (T), a steroidal alkaloid abundantly found in green tomatoes for maximizing intracellular delivery of lipoplexes. We doped Tomatidine into liposomes of cationic lipid with amide linker (A) from our lipid library. Six liposomal formulations (AT) of Lipid A (1 mM) with varying concentrations of Tomatidine (0–1 mM) were prepared and evaluated for their transfection efficacies. Owing to its signature characteristic of cell membrane permeability, Tomatidine modulated endocytosis process, enhanced the intracellular delivery of the lipoplexes, and in turn increased the transfection efficacy of cationic liposomes. Our findings provide ‘proof of concept’ for enhancing transfections in gene delivery applications with Tomatidine in cationic liposomal formulations. These findings can be further applied in lipid mediated gene therapy and drug delivery applications.

UR - https://www.sciencedirect.com/science/article/pii/S0005273618301780 ER -