Synthesis of Cationic Di-block Glycopolypeptides for Dendritic Cell-based Gene Transfection

Mukhopadhyay, Rounak (2025) Synthesis of Cationic Di-block Glycopolypeptides for Dendritic Cell-based Gene Transfection. Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS Dissertation of Rounak Mukhopadhyay (20MS189)) 20MS189_Thesis_file.pdf - Submitted Version Restricted to Repository staff only Download (2MB)

Abstract

Gene therapy has emerged as a transformative approach for treating genetic disorders by delivering therapeutic nucleic acids into target cells. However, efficient gene delivery—particularly to immune cells like dendritic cells (DCs)—remains a significant challenge due to issues such as cytotoxicity, immune activation, poor endosomal escape, and low targeting specificity. While viral vectors have demonstrated high transfection efficiencies, their clinical utility is hindered by immunogenicity, insertional mutagenesis, and production complexity. In this context, non-viral vectors, especially synthetic polypeptides, offer safer, more tunable platforms for gene transfection owing to their biodegradability, modularity, and biomimetic properties. This thesis presents the rational design and development of two cationic di-block glycopolypeptides tailored for gene delivery to DCs. The first design incorporates a guanidinium-functionalized glutamic acid block (Gua-TEG-Glu) for strong DNA binding and α-helical stability, paired with a lysine-shikimic acid block (LSA) for mannose receptor-mediated targeting. The second design features a lysine-histidine block (Lys-His) to enhance endosomal escape through the proton sponge effect, combined with the same LSA targeting module. Both constructs maintain α-helical conformation for membrane interaction and multivalent presentation of functional groups, improving transfection efficacy while reducing immunotoxicity. The modular architecture of these glycopolypeptides allows for the separation of functions—nucleic acid condensation, targeting, and intracellular trafficking—mimicking natural delivery systems such as viral capsids. Compared to previous designs, such as homoarginine-based systems which suffered from poor helicity and buffering capacity, these constructs offer improved biophysical and biological properties. This work provides a promising foundation for non-viral gene delivery systems targeting DCs, potentially advancing gene-based immunotherapies for cancer, autoimmune, and infectious diseases.

Actions (login required)

View Item