Compositions and methods for the treatment of hemoglobinopathies

Patent metadata

Field	Value
Publication number	US-10619140-B2
Application number	US-201715461091-A
Country	US
Kind code	B2
Filing date	Mar 16, 2017
Priority date	Feb 24, 2012
Publication date	Apr 14, 2020
Grant date	Apr 14, 2020

Abstract

Provided are compositions and methods for the treatment of hemoglobinopathies such as thalassemias and sickle cell disease. Compositions and methods include one or more endonuclease(s) or endonuclease fusion protein(s), including one or more homing endonuclease(s) and/or homing endonuclease fusion protein(s) and/or CRISPR endonuclease(s) and/or CRISPR endonuclease fusion protein(s): (a) to disrupt a Bcl11a coding region; (b) to disrupt a Bcl11a gene regulatory region; (c) to modify an adult human β-globin locus; (d) to disrupt a HbF silencing DNA regulatory element or pathway, such as a Bcl11a-regulated HbF silencing region; (e) to mutate one or more γ-globin gene promoter(s) to achieve increased expression of a γ-globin gene; (f) to mutate one or more δ-globin gene promoter(s) to achieve increased expression of a δ-globin gene; and/or (g) to correct one or more β-globin gene mutation(s).

First claim

What is claimed is:

1. An I-OnuI homing endonuclease (HE) variant that binds to a target site in a fetal hemoglobin (HbF) silencing region set forth in SEQ ID NO: 2,
wherein the I-OnuI HE variant comprises one or more amino acid substitutions at positions L26, R28, R30, N32, S40, E42, G44, Q46, A70, S72, S78, K80, and T82 set forth in SEQ ID NO: 15;
wherein the I-OnuI HE variant comprises one or more amino acid substitutions at positions F182, N184, 1186, S190, K191, Q197, V199, S201, K225, K227, D236, V238, and T240 set forth in SEQ ID NO: 15; and
wherein the I-OnuI HE variant is fused to a TALE DNA binding domain that binds a nucleotide sequence within the HbF silencing region and adjacent to the target site.

2. The I-OnuI HE variant of claim 1 , wherein the I-OnuI HE variant further comprises a TREX2 nuclease domain.

3. A polynucleotide encoding the I-OnuI variant of claim 1 or claim 2 .

4. A vector comprising the polynucleotide of claim 3 .

5. The vector system of claim 4 , wherein the vector is selected from the group consisting of an AAV6, a modified adenovirus vector, an integration-deficient lentiviral vector (IDLV), and an integration-deficient foamyviral vector (IDFV).

6. A composition comprising the I-OnuI variant of claim 1 or claim 2 .

7. A composition comprising the polynucleotide of claim 3 .

8. A cell comprising the I-OnuI variant of claim 1 or claim 2 .

9. A cell comprising the polynucleotide of claim 3 .

Assignees

• Hutchinson Fred Cancer Res

Inventors

• Takeuchi Ryo
• Groudine Mark T
• Stoddard Barry L
• Bender Michael A

Classifications

• C12N9/16 Primary

  acting on ester bonds (3.1) · CPC title

• C07K2319/80

  containing a DNA binding domain, e.g. Lacl or Tet-repressor · CPC title

• A61P7/00

  Drugs for disorders of the blood or the extracellular fluid · CPC title

• A61P7/06

  Antianaemics · CPC title

• A61K48/00

  Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy · CPC title