Recombinant therapeutic cytokines (therakines) for the treatment of cancer are provided. The recombinant therakines include a truncated region of MDA-7/TL-24 (M4) not normally found in nature that has anti-cancer activity and a secretory signal which causes secretion of the therakine from plasmid/virus transduced normal and cancer cells and interaction of the therakine to MDA-7/IL-24 receptors on adjacent, neighboring and distant cancer cells. Therakine interaction results in bystander killing of the target cancer cell as well as adjacent, neighboring and distant cancer cells.

The present application relates to methods of producing exosomes. The application also provides a method for preparing a protein composition comprising culturing an exosome-producing cell expressing a Nef-fusion protein comprising a Nef-derived peptide fused to a protein of interest; isolating exosomes from the exosome-producing cell culture; and purifying the protein of interest from the isolated exosomes. The application further discloses compositions that comprise exosomes containing the Nef-fusion protein, as well as methods of using the Nef-fusion protein and exosomes containing the Nef-fusion protein.

Provided are polynucleotides and viral vectors, particularly, Alphavirus vectors such as Sindbis viral vectors, which encode an immune checkpoint protein, or a ligand binding portion of the checkpoint protein, or an immune checkpoint protein or ligand binding portion thereof fused to one or more immunoglobulin (Ig) domains, e.g., an Ig hinge region and an Ig heavy chain constant domain. Methods of treating a mammalian subject having a cancer or tumor are provided, in which the viral vectors, e.g., a Sindbis virus vector, encoding the immune checkpoint protein, a ligand binding portion thereof, or a checkpoint protein fusion protein as described, are administered to the subject, resulting in an anti-cancer or anti-tumor immune response, significant reduction in tumor growth in the treated subject and increased survivability.

Proteins, nucleic acids encoding the proteins, compositions comprising the proteins, and methods are provided. The proteins have the ability to be self-targeted to ICAM-1 and, if desired, enzymatically-released at acidic pH. The ICAM-1-targeting peptides are provided as single copies or multiples repeats, and can be separated by linkers from the enzyme segment, from which the ICAM-1 targeting peptides can be released, if desired, at acidic pH. These fusion proteins enhance the activity of the enzyme segment within or liberated from the fusion protein, and provide increased recognition and targeting of diseased organs, transport from the bloodstream across the endothelium into said diseased organ, and intracellular uptake and lysosomal trafficking by cells in them, both in peripheral tissues and the central nervous system. Representative nucleotide and amino acid sequences of these fusion proteins, as well as in vitro, cellular, and in vivo animal data are provided. The described proteins can be used as a protein therapy, a gene therapy, or an implanted cell therapy.

Provided herein are fusion constructs including a biologically active agent such as an antibody or derivative thereof which is functionally linked with a functional moiety such as an LC3 protein or portion thereof. Also provided are compositions and exosomes including such constructs, as well as methods for preparing exosomes containing such constructs, and methods for delivering such constructs to cells. Methods and uses of such constructs and exosomes for treating diseases or disorders are also provided, in which the constructs trigger autophagy of disease-related cellular or cytoplasmic targets such as misfolded/aggregated proteins in neurodegenerative diseases, for example.

The present application relates to methods of producing exosomes. The application also provides a method for preparing a protein composition comprising culturing an exosome-producing cell expressing a Nef-fusion protein comprising a Nef-derived peptide fused to a protein of interest; isolating exosomes from the exosome-producing cell culture; and purifying the protein of interest from the isolated exosomes. The application further discloses compositions that comprise exosomes containing the Nef-fusion protein, as well as methods of using the Nef-fusion protein and exosomes containing the Nef-fusion protein.

The present invention relates to a novel enhancer of protein production in host cells. It discloses a vector for expressing recombinant proteins in these cells, comprising a nucleotide sequence encoding a) a secretion peptidic signal, b) a 6-methylguanine-DNA-methyltransferase enzyme (MGMT, EC 2.1.1.63), a mutant or a catalytic domain thereof, and c) a recombinant protein. Said MGMT enzyme is preferably the so-called SNAP protein.

Recombinant therapeutic cytokines (therakines) for the treatment of cancer are provided. The recombinant therakines include a truncated region of MDA-7/TL-24 (M4) not normally found in nature that has anti-cancer activity and a secretory signal which causes secretion of the therakine from plasmid/virus transduced normal and cancer cells and interaction of the therakine to MDA-7/IL-24 receptors on adjacent, neighboring and distant cancer cells. Therakine interaction results in bystander killing of the target cancer cell as well as adjacent, neighboring and distant cancer cells.

Fluorescent and non-fluorescent fusion tags derived from green fluorescent protein (GFP). The fusion tags harbor substitutions with respect to GFP that increase expression of target proteins to which the fusion tags are fused; increase solubility; prevent dimerization and oligomerization, particularly in oxidizing environments such as the endoplasmic reticulum (ER); and, in some cases, enhance fluorescence of the fusion tag itself. The substitutions include various combinations of substitutions at select cysteine residues in GFP, substitutions in or near the GFP chromophore, and/or other substitutions. The fusion tags can be used for increasing expression of target proteins for mass production thereof or as a fluorescent tag.

The current disclosure relates to pro-apoptotic molecules with a B-cell lymphoma 2 (BCL-2) homology 3 (BH3) effector domain. The current disclosure furthermore relates to pro-apoptotic constructs wherein the pro-apoptotic molecules are linked to a granule-localizing domain. The pro-apoptotic construct may be transferred from an effector cell to a target cell to induce apoptosis. The current disclosure also relates to the nucleic acid molecules encoding the pro-apoptotic proteins and the uses thereof in a medical therapy such as cancer therapy, including chimeric antigen receptor cell therapy and the like.