Preparation and In Vitro Evaluation of RITUXfab-Decorated Lipoplexes to Improve Delivery of siRNA Targeting C1858T PTPN22 Variant in B Lymphocytes

• Published in: International journal of molecular sciences 2021-12, Vol.23 (1), p.408
• Main Authors: Arena, Andrea, Belcastro, Eugenia, Accardo, Antonella, Sandomenico, Annamaria, Pagliarosi, Olivia, Rosa, Elisabetta, Petrini, Stefania, Conti, Libenzio Adrian, Giorda, Ezio, Corsetti, Tiziana, Schiaffini, Riccardo, Morelli, Giancarlo, Fierabracci, Alessandra
• Format: Article
• Language: English
• Subjects: Adaptive immunity; Amino Acid Sequence; Antigens; autoimmune disease; Autoimmune diseases; B-Lymphocytes - drug effects; B-Lymphocytes - immunology; Cell differentiation; Cell Line; Cell proliferation; Chromatography; Circular Dichroism; CpG islands; Customization; Diabetes; Diabetes mellitus (insulin dependent); Differentiation (biology); Dynamic Light Scattering; Endocrine disorders; functionalized lipoplexes; Gene polymorphism; Gene Transfer Techniques; Genetic diversity; Genomes; Health risk assessment; Hormone replacement therapy; Humans; Immunoglobulin Fab Fragments - immunology; Immunoglobulin M; Immunotherapy; Lipids - chemistry; Liposomes; Lymphocyte Activation - immunology; Lymphocytes; Lymphocytes B; Monoclonal antibodies; Mutation - genetics; Pathogenesis; Peripheral blood mononuclear cells; Phenotype; Phosphatase; Polymorphism; Polypeptides; Protein Tyrosine Phosphatase, Non-Receptor Type 22 - genetics; Protein-tyrosine-phosphatase; Proteolysis - drug effects; Rituximab; Rituximab - chemistry; Rituximab - immunology; Rituximab - pharmacology; RNA, Small Interfering - administration & dosage; siRNA; T1D; Thyroid diseases; Thyroiditis; Tumor necrosis factor-TNF; Tyrosine; variant PTPN22
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms23010408

Autoimmune endocrine disorders, such as type 1 diabetes (T1D) and thyroiditis, at present are treated with only hormone replacement therapy. This emphasizes the need to identify personalized effective immunotherapeutic strategies targeting T and B lymphocytes. Among the genetic variants associated with several autoimmune disorders, the C1858T polymorphism of the protein tyrosine phosphatase non-receptor type 22 ( ) gene, encoding for Lyp variant R620W, affects the innate and adaptive immunity. We previously exploited a novel personalized immunotherapeutic approach based on siRNA delivered by liposomes (lipoplexes) that selectively inhibit variant allele expression. In this manuscript, we improved lipoplexes carrying siRNA for variant C1858T by functionalizing them with Fab of Rituximab antibody (Ritux -Lipoplex) to specifically target B lymphocytes in autoimmune conditions, such as T1D. Ritux -Lipoplexes specifically bind to B lymphocytes of the human Raji cell line and of human PBMC of healthy donors. Ritux -Lipoplexes have impact on the function of B lymphocytes of T1D patients upon CpG stimulation showing a higher inhibitory effect on total cell proliferation and IgM+ plasma cell differentiation than the not functionalized ones. These results might open new pathways of applicability of Ritux -Lipoplexes, such as personalized immunotherapy, to other autoimmune disorders, where B lymphocytes are the prevalent pathogenic immunocytes.