Compositions for facilitating membrane fusion and uses thereof

Abstract

In some aspects, fusosome compositions and methods are described herein that comprise membrane enclosed preparations, comprising a fusogen. In some embodiments, the fusosome can fuse to the target cell, thereby delivering complex biologic agents to the target cell cytoplasm.

1 . A fusosome composition comprising a plurality of fusosomes derived from a source cell, wherein the fusosomes of the plurality comprise:
 (a) a lipid bilayer,   (b) a lumen comprising cytosol from the source cell, wherein the lumen is surrounded by the lipid bilayer,   (c) an exogenous or overexpressed fusogen disposed in the lipid bilayer, wherein the fusogen comprises a protein selected from the group consisting of Nipah virus F and G proteins, measles virus F and H proteins, tupaia paramyxovirus F and H proteins, paramyxovirus F and G proteins or F and H proteins or F and HN proteins, Hendra virus F and G proteins, Henipavirus F and G proteins, Morbilivirus F and H proteins, respirovirus F and HN protein, a Sendai virus F and HN protein, rubulavirus F and HN proteins, or avulavirus F and HN proteins, a derivative thereof, and any combination thereof,   (d) a cargo; and   wherein the fusosomes do not comprise a nucleus;   wherein the amount of viral capsid protein in the fusosome composition is less than 1% of total protein.   
     
     
         2 .- 6 . (canceled) 
     
     
         7 . The fusosome composition of  claim 1 , wherein the fusosomes of the plurality further comprise a targeting moiety. 
     
     
         8 . The fusosome composition of  claim 7 , wherein the targeting moiety is comprised by the fusogen or is comprised by a separate molecule. 
     
     
         9 .- 13 . (canceled) 
     
     
         14 . The fusosome composition of  claim 7 , wherein the targeting moiety is specific for a cell surface marker on the target cell. 
     
     
         15 . The fusosome composition of  claim 14 , wherein the cell surface marker is a cell surface marker of a skin cell, a cardiomyocyte, a hepatocyte, an intestinal cell, a pancreatic cell, a brain cell, a prostate cell, a lung cell, a colon cell, or a bone marrow cell. 
     
     
         16 .- 20 . (canceled) 
     
     
         21 . The fusosome composition of  claim 1 , wherein the fusosomes of the plurality comprise exosomes, microvesicles, or a combination thereof. 
     
     
         22 . The fusosome composition of  claim 1 , wherein the fusosomes of the plurality have an average size of at least 50 nm, 100 nm, 200 nm, 500 nm, 1000 nm, 1200 nm, 1400 nm, or 1500 nm in diameter. 
     
     
         23 . (canceled) 
     
     
         24 . The fusosome composition of  claim 1 , wherein the source cell is selected from the group consisting of a neutrophil, a HEK293 cell, a granulocyte, a mesenchymal stem cell, a bone marrow stem cell, an induced pluripotent stem cell, an embryonic stem cell, a myeloblast, a myoblast, a hepatocyte, or a neuron. 
     
     
         25 . The fusosome composition of  claim 1 , wherein the fusosomes of the plurality comprise cell with partial or complete nuclear inactivation. 
     
     
         26 . The fusosome composition of  claim 1 , wherein the fusosomes of the plurality comprise an enucleated cell. 
     
     
         27 .- 31 . (canceled) 
     
     
         32 . The fusosome composition of  claim 1 , wherein the fusogen comprises a sequence chosen from a Nipah virus protein F, a measles virus F protein, a tupaia paramyxovirus F protein, a paramyxovirus F protein, a Hendra virus F protein, a Henipavirus F protein, a Morbilivirus F protein, a respirovirus F protein, a Sendai virus F protein, a rubulavirus F protein, or an avulavirus F protein, or a derivative thereof. 
     
     
         33 . (canceled) 
     
     
         34 . The fusosome composition of  claim 1 , wherein the fusosomes in the plurality further comprises a Nipah virus protein G, a measles protein H, a tupaia paramyxovirus H protein, a paramyxovirus G protein, a paramyxovirus H protein, a paramyxovirus HN protein, a Morbilivirus H protein, a respirovirus HN protein, a sendai HN protein, a rubulavirus HN protein, an avulavirus HN protein, or a derivative thereof. 
     
     
         35 . (canceled) 
     
     
         36 . The fusosome composition of  claim 1 , wherein the cargo comprises or encodes:
 (a) an exogenous protein or an exogenous nucleic acid,   (b) a cytosolic protein or a membrane protein: or   (c) a therapeutic agent.   
     
     
         37 .- 44 . (canceled) 
     
     
         45 . A pharmaceutical composition comprising the fusosome composition of  claim 1  and pharmaceutically acceptable carrier. 
     
     
         46 . The pharmaceutical composition of  claim 45 , wherein the cargo comprises a therapeutic agent. 
     
     
         47 . A method of delivering a cargo to a subject, comprising administering to the subject the pharmaceutical composition of  claim 46 , wherein the fusosome composition is administered in an amount and/or at a time such that the cargo is delivered. 
     
     
         48 . (canceled) 
     
     
         49 . (canceled) 
     
     
         50 . A fusosome composition comprising a plurality of fusosomes derived from a source cell, wherein the fusosomes of the plurality comprise:
 (a) a lipid bilayer,   (b) a lumen comprising cytosol from the source cell, wherein the lumen is surrounded by the lipid bilayer,   (c) an exogenous or overexpressed fusogen, wherein the fusogen is disposed in the lipid bilayer and wherein the fusogen comprises a protein selected from the group consisting of Nipah virus F and G proteins, measles virus F and H proteins, tupaia paramyxovirus F and H proteins, paramyxovirus F and G proteins or F and H proteins or F and HN proteins, Hendra virus F and G proteins, Henipavirus F and G proteins, Morbilivirus F and H proteins, respirovirus F and HN protein, a Sendai virus F and HN protein, rubulavirus F and HN proteins, or avulavirus F and HN proteins, a derivative thereof, and any combination thereof, and   (d) a cargo;   
       wherein the fusosomes do not comprise a nucleus. 
     
     
         51 .- 53 . (canceled) 
     
     
         54 . The fusosome composition of  claim 1 , which does not comprise a viral capsid protein. 
     
     
         55 . A pharmaceutical composition comprising the fusosome composition of  claim 50  and pharmaceutically acceptable carrier. 
     
     
         56 . The pharmaceutical composition of  claim 55 , wherein the cargo comprises a therapeutic agent. 
     
     
         57 . A method of delivering a cargo to a subject, comprising administering to the subject the pharmaceutical composition of  claim 56 , wherein the fusosome composition is administered in an amount and/or at a time such that the cargo is delivered. 
     
     
         58 . The fusosome composition of  claim 8 , wherein the targeting moiety is specific for a cell surface marker on the target cell