Patent WO-2023279121-A9 (WO)
Inventor: SWARTZ ROGER
IPC: C12N15/867
Filed: 2022-07-07
Abstract: The present invention contemplates mRNA, episomal and retroviral genomic gene therapy based short-term, intermediate or long-term up to multidecade vaccine, immunization, immune protection or cancer - that can also be administered as a retroviral genomic gene therapy both in vivo and ex vivo - method to provide epithelial and hematological protection to humans to protect against cancer especially carcinomas, pandemic and non-pandemic viruses, bacterial infections, allergens or the cause of allergic reactions, systemic pathological conditions, cancer and anti-biowarfare agents (e.g. natural and unnatural viruses and toxins) where mucosal immunity and for some diseases hematological immunity is achieved through mRNA, episomal or genomic integrated lentiviral and gammaretroviral vector expression of dimeric immunoglobulin A1 (dlgA1), dimeric immunoglobulin A2 (dlgA2) and engineered variants. Additionally, in some embodiments a method to agglutinate cancers including carcinomas and hematological cancers to prevent metastasis with polymeric immunoglobulin A, dlgA and engineered variants such as but not limited to engineering the hinge region of the immunoglobulin. The present invention provides vector constructs to express dlgA and polymeric immunoglobulins A with therapeutically relevant activity (e.g. such as based on complementarity determining Regions (CDRs) or V-regions) against its target in human. Antibody V-regions or CDR regions may be discovered in human. Alternatively, combinatorial libraries of single chain variable fragments (scFV) will generated from human B-cells or other animal B-cells where phage display technology and mutagenesis can be used to identify potent VH and VL immunoglobulin fragments that can be incorporated into full-length or reduced length immunoglobulin heavy and light chains. Further, mice or other animals with humanized immune systems or other non-human vertebrate such as rabbits or hamsters capable of producing fully human antibodies from exposure to antigen may be used to identify potent immunoglobulins or potent immunoglobulin binding regions or potent immunoglobulin complementarity determining regions (CDRs) for fusion with human antibodies giving rise to chimeric antibodies. The identified immunoglobulins from these methods will optionally be further optimized through mutagenesis techniques and will be expressed in the recipient via mRNA, via an episome or via retroviral insertion into their genomic DNA of the cells of interest to be expressed via intramuscular administration, intravenous administration, endoscopy based administration to the lamina propria of the stomach and/or small intestine or even the lung, via ingestion or administration proximal to lymph nodes or as an ex vivo administration into any of B-cells, T-cells, Natural Killer (NK) Cells and other immune cell types. Preferred cells to target to receive the vector include muscle cells, liver cells especially hepatocytes and B-cells including memory B-cells, Germinal Center B-cells, memory plasma B-cells (also referred to as a long-lived plasma cell), naive B-cells, NK cells, T-cells, including chimeric antigen receptor T-cells (CAR T-cells) as well as any CAR engineered immune cell. Additionally, the vector may encode for both the CAR and the polymeric and dimeric immunoglobulin in a single vector construct. When designing an mRNA, AAV viral vector, adenovirus vector, integration deficient lentivirus retroviral vector or gammaretroviral vector, integration deficient lentivirus retroviral vector or gammaretroviral vector, encoding for dlgA1, dlgA2 or polymeric immunoglobulin A a single DNA-based vector or one or more mRNA vectors will code for the entire immunoglobulin and J Chain (Joining Chain) expression for dlgA1 or dlgA2, where expression may occur with a single start codon and stop codon for each transgene and in some embodiments a second start codon for J Chain expression. The use of a single start and stop codon is enabled by placing in the 5 to 3 direction a furin cleavage site concomitantly followed by a 2A self-processing peptide or furin cleavage site between each gene of any number of consecutive transgenes as a single open reading frame.