Genetically engineered hematopoietic stem cell as a platform for systemic protein expression

1. A genetically modified hematopoietic stem cell comprising, in at least one globin gene comprised in the genome thereof, at least one transgene encoding a therapeutic protein or a therapeutic ribonucleic acid, the transgene being placed under control of the endogenous promoter of the at least one globin gene,

wherein the at least one transgene encoding a therapeutic protein or a therapeutic ribonucleic acid is comprised in the 5′ region, in the 3′ untranslated region (3′ UTR) and/or in an intron of the at least one globin gene,

wherein the at least one globin gene is selected from the group consisting of the alpha 1 globin gene and the alpha 2 globin gene.

2. The genetically modified hematopoietic stem cell according to claim 1 , wherein the at least one transgene encoding a therapeutic protein or a therapeutic ribonucleic acid is comprised in the 5′ region and/or in the second intron (IVS2) of the at least one globin gene.

3. The genetically modified hematopoietic stem cell according to claim 1 , wherein the at least one transgene encoding a therapeutic protein or a therapeutic ribonucleic acid is comprised in the 5′ untranslated region (5′UTR) and/or in the proximal promoter and/or in the second intron (IVS2) of the at least one globin gene.

4. The genetically modified hematopoietic stem cell according to claim 1 , wherein

the at least one globin gene is the alpha 1 globin gene and/or the alpha 2 globin gene;

and the at least one transgene encoding a therapeutic protein or a therapeutic ribonucleic acid is comprised in the 5′ untranslated region (5′UTR) or in an intron, of the at least one globin gene.

5. The genetically modified hematopoietic stem cell according to claim 1 , wherein the encoded therapeutic protein is selected from the group consisting of cytokines; hormones; chemokines; antibodies; anti-angiogenic factors; enzymes for replacement therapy; insulin; G-CSF; GM-CSF; hPG-CSF; M-CSF; blood clotting factors; transmembrane proteins; lysosomal enzymes; any protein that can be engineered to be secreted and eventually taken up by non-modified cells, and combinations thereof.

6. A blood cell originating from a genetically modified hematopoietic stem cell according to claim 1 .

7. A pharmaceutical composition comprising at least one genetically modified hematopoietic stem cell according to claim 1 and/or at least one blood cell originating from the at least one genetically modified hematopoietic stem cell, and a pharmaceutically acceptable medium.

8. A method for the ex vivo or in vitro preparation, of the genetically modified hematopoietic stem cell according to claim 1 , comprising the steps of:

(i) providing to a hematopoietic stem cell a site-directed genetic engineering system by:

(a) providing to the hematopoietic stem cell (1) at least one guide nucleic acid binding to a selected target site or (2) a guide peptide-containing endonuclease binding to a selected target site, the selected target site being located in an endogenous globin-encoding gene comprised in the genome of the hematopoietic stem cell, the endogenous globin-encoding gene being selected from the group consisting of the alpha 1 globin gene and the alpha 2 globin gene;

(b) after the at least one guide nucleic acid has been provided at step a), further providing to the hematopoietic stem cell at least one endonuclease devoid of target site specificity; and

(c) further providing to the hematopoietic stem cell a transgene that encodes at least one therapeutic protein or at least one therapeutic ribonucleic acid; and

(ii) culturing the hematopoietic stem cell obtained at step (i) such that the transgene is introduced at the said selected target site in the genome of the hematopoietic stem cell.

9. The method according to claim 8 , wherein the method further comprises the steps of:

(i) providing to the hematopoietic stem cell a site-directed genetic engineering system by:

(a) providing to the hematopoietic stem cell at least one guide nucleic acid binding to a selected target site, the target site being located in the endogenous globin-encoding gene comprised in the genome of the hematopoietic stem cell;

(b) further providing to the hematopoietic stem cell at least one endonuclease devoid of target site specificity; and

(c) further providing to the hematopoietic stem cell a transgene that encodes at least one therapeutic protein or at least one therapeutic ribonucleic acid; and

(ii) culturing the hematopoietic stem cell obtained at the end of step (i) such that the transgene is introduced at the selected target site in the genome of the hematopoietic stem cell.

10. The method according to claim 8 , wherein the at least one endonuclease devoid of target site specificity is a Clustered regularly interspaced short palindromic repeats (CRISPR) associated nuclease.

11. The method according to claim 8 , wherein the at least one guide nucleic acid is a guide RNA which recognizes a target site in the 5′ region and/or in an intron and/or in the proximal promoter and/or in an intron of the at least one globin gene comprised in the genome of the hematopoietic stem cell.

12. A method for the treatment of

a disease selected from the group consisting of autoimmune diseases, viral infections and tumors; and/or

a disease caused by the lack of a protein or by the presence of an aberrant non-functional protein in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of the genetically modified hematopoietic stem cell according to claim 1 , a blood cell originating from the genetically modified hematopoietic stem cell, or a pharmaceutical composition comprising the genetically modified hematopoietic stem cell or the blood cell.

13. A method for inducing immune tolerance in an individual in need thereof, comprising administering to the individual a hematopoietic stem cell according to claim 1 , a blood cell originating from the hematopoietic stem cell, or a pharmaceutical composition comprising the genetically modified hematopoietic stem cell or the blood cell.