# US 10,865,379 B2 · Methods and compositions for promoting survival and proliferation of endothelial cells and stimulating angiogenesis > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,865,379 B2 - Document ID: US10865379B2 - Status: Granted - Kind: B2 - Application number: 15/906,408 - Filed: 2018-02-27 - Published: 2018-09-13 - Granted: 2020-12-15 - Earliest claimed: 2007-01-19 - Assignees: Cornell Research Foundation, Inc.; Sloan-Kettering Institute for Cancer Research - Applicants: Cornell Research Foundation, Inc.; Sloan-Kettering Institute for Cancer Research - Inventors: Shahin Rafii; Fan Zhang; Marco Seandel - Web page: https://patentgrep.com/patent/US10865379B2 ## CPC - C12N 5/0606: {Pluripotent embryonic cells, e.g. embryonic stem cells [ES] (embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696)} - C07K 14/005: from viruses - C12N 5/0647: {Haematopoietic stem cells; Uncommitted or multipotent progenitors} - C12N 5/069: {Vascular Endothelial cells} - C12N 5/0693: {Tumour cells; Cancer cells} - C12N 15/1131: {against viruses} - C12N 15/1136: {against growth factors, growth regulators, cytokines, lymphokines or hormones} - C12N 2310/14: interfering nucleic acids [NA] - C12N 2501/998: Proteins not provided for elsewhere - C12N 2710/10322: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - C12N 2740/10043: viral genome or elements thereof as genetic vector - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract The present invention relates to adenovirus E4ORF1 gene and to endothelial cells engineered to express the E4ORF1 gene. The present invention also relates to uses of the E4ORF1 gene, and cells expressing the E4ORF1 gene, and to compositions comprising the E4ORF1 gene, or comprising cells expressing the E4ORF1 gene. ## Claims ### Claim 1 A composition comprising a population endothelial cells, wherein the endothelial cells (a) comprise a recombinant nucleic acid sequence that comprises an E4ORF1 coding sequence under the control of a promoter, and (b) do not comprise nucleic acid sequences that comprise the entire adenovirus E4 region. ### Claim 2 (depends on 1) The composition of claim 1, wherein the recombinant nucleic acid sequence is comprised within a lentiviral vector. ### Claim 3 (depends on 1) The composition of claim 1, wherein the recombinant nucleic acid sequence is comprised within an adenoviral vector. ### Claim 4 (depends on 1) The composition of claim 1, wherein the recombinant nucleic acid sequence further comprises an antibiotic resistance gene. ### Claim 5 (depends on 1) The composition of claim 1, wherein the recombinant nucleic acid sequence further comprises a reporter gene or a sequence encoding an expression tag. ### Claim 6 (depends on 5) The composition of claim 5, wherein the recombinant nucleic acid sequence encodes green fluorescent protein (GFP). ### Claim 7 (depends on 1) The composition of claim 1, wherein the promoter is selected from the group consisting of a CMV promoter, an SV40 promoter, an RSV promoter, an HIV-Ltr promoter, and an MML promoter. ### Claim 8 (depends on 1) The composition of claim 1, wherein the promoter is an adenovirus promoter. ### Claim 9 (depends on 1) The composition of claim 1, wherein the promoter is an inducible promoter. ### Claim 10 (depends on 1) The composition of claim 1, wherein the E4ORF1 coding sequence is a human adenovirus type 5 coding sequence. ### Claim 11 (depends on 1) The composition of claim 1, wherein the E4ORF1 coding sequence comprises SEQ ID NO. 1. ### Claim 12 (depends on 1) The composition of claim 1, wherein the E4ORF1 coding sequence is an E4ORF1 coding sequence from human adenovirus 1, human adenovirus 2, human adenovirus 3, human adenovirus 7, human adenovirus 9, human adenovirus 11, human adenovirus 12, human adenovirus 14, human adenovirus 34, human adenovirus 35, human adenovirus 46, human adenovirus 50, or human adenovirus 52. ### Claim 13 (depends on 1) The composition of claim 1, where the endothelial cells are selected from the group consisting of: testis endothelial cells, lung endothelial cells, lymphatic endothelial cells, cord blood endothelial cells, umbilical vein endothelial cells, heart endothelial cells, skin endothelial cells, liver endothelial cells, brain endothelial cells, bone endothelial cells, and pancreatic endothelial cells. ### Claim 14 (depends on 1) The composition of claim 1, further comprising a population of cancer cells, stem cells, or progenitor cells. ### Claim 15 (depends on 14) The composition of claim 14, wherein the stem cells or progenitor cells are selected from the group consisting of embryonic stem cells, hematopoietic stem or progenitor cells, neural stem or progenitor cells, skin stem or progenitor cells, spermatogonial stem or progenitor cells, and gut stem or progenitor cells. ### Claim 16 (depends on 1) The composition of claim 1, further comprising a population of hematopoietic stem or progenitor cells. ### Claim 17 (depends on 1) The composition of claim 1, further comprising Matrigel. ### Claim 18 (depends on 1) The composition of claim 1, further comprising a physiological saline solution or a carrier solution suitable for administration to a living subject. ### Claim 19 (depends on 1) A method of inducing angiogenesis in vivo, the method comprising administering a composition according to claim 1 to a subject in need thereof. ### Claim 20 (depends on 1) A method of inducing wound healing in vivo, the method comprising administering a composition according to claim 1 to a subject in need thereof. ### Claim 21 A method for maintaining or expanding cancer cells, stem cells or progenitor cells in vitro, the method comprising: a. contacting cancer cells, stem cells or progenitor cells in vitro with conditioned medium, wherein the condition medium is derived from a culture of E4ORF1-expressing endothelial cells that do not contain or express nucleic acid sequences that comprise the entire adenovirus E4 region, thereby maintaining or expanding the cancer cells, stem cells or progenitor cells in vitro. ### Claim 22 A method of maintaining or expanding cancer cells, stem cells or progenitor cells in vitro, the method comprising: b. culturing a population of cancer cells, stem cells or progenitor cells in vitro in a culture vessel that comprises a population of E4ORF1-expressing endothelial cells, wherein the endothelial cells do not contain or express nucleic acid sequences that comprise the entire adenovirus E4 region, thereby maintaining or expanding the cancer cells, stem cells or progenitor cells in vitro.