Factor viii mutation repair and tolerance induction
AssigneesHaplomics, Inc., The Regents of the University of California, The United States Government AS Represented By the Deparment of Veterans Affairs
InventorsVincent LATERZA, Tommy E. HOWARD
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Abstract
Methods of treating hemophilia A in a subject with an F8 gene mutation, wherein the F8 gene is repaired and the resultant repaired gene, upon expression, confers improved coagulation functionality to the encoded FVIII protein of the subject compared to the non-repaired F8 gene. The invention also includes methods of inducing immune tolerance to a FVIII replacement product ((r)FVIII) in a subject having a FVIII deficiency, wherein the F8 gene mutation is repaired and the repaired gene, upon expression, provides for the induction of immune tolerance to an administered replacement FVIII protein product. The invention also includes isolated nucleic acids, vectors, recombinant viruses, cells, and pharmaceutical compositions to repair the F8 gene.
Claims20
A method of treating hemophilia A in a subject comprising:
- introducing into a cell of the subject one or more isolated nucleic acids encoding a nuclease that targets a portion of the F8 gene containing a mutation that causes hemophilia A, wherein the nuclease creates a double stranded break in the F8 gene;
- and an isolated nucleic acid comprising a donor sequence comprising (i) a nucleic acid encoding a truncated FVIII polypeptide or (ii) a native F8 3′ splice acceptor site operably linked to a nucleic acid encoding a truncated FVIII polypeptide,
- wherein the nucleic acid comprising the (i) nucleic acid encoding a truncated FVIII polypeptide or (ii) native F8 3′ splice acceptor site operably linked to a nucleic acid encoding a truncated FVIII polypeptide is flanked by nucleic acid sequences homologous to the nucleic acid sequences upstream and downstream of the double stranded break in the DNA, and
- wherein the resultant repaired gene, upon expression, confers improved coagulation functionality to the encoded FVIII protein of the subject compared to the non-repaired F8 gene.
A method of inducing immune tolerance to a FVIII replacement product ((r)FVIII) in a subject having a FVIII deficiency and who will be administered, is being administered, or has been administered a (r)FVIII product comprising:
- introducing into a cell of the subject one or more nucleic acids encoding a nuclease that targets a portion of the F8 gene containing a mutation that causes hemophilia A,
- wherein the nuclease creates a double stranded break in the F8 gene;
- and an isolated nucleic acid comprising a donor sequence comprising (i) a nucleic acid encoding a truncated FVIII polypeptide or (ii) a native F8 3′ splice acceptor site operably linked to a nucleic acid encoding a truncated FVIII polypeptide,
- wherein the nucleic acid comprising the (i) nucleic acid encoding a truncated FVIII polypeptide or (ii) native F8 3′ splice acceptor site operably linked to a nucleic acid encoding a truncated FVIII polypeptide is flanked by nucleic acid sequences homologous to the nucleic acid sequences upstream and downstream of the double stranded break in the DNA, and
- wherein the repaired gene, upon expression, provides for the induction of immune tolerance to an administered replacement FVIII protein product.
An isolated nucleic acid encoding a truncated Factor VIII polypeptide consisting essentially of the amino acid sequence encoded by exons 23, 24, 25 and 26 of human Factor VIII.
-34. (canceled)
Dependencies are inferred from the claim text.