Compositions and methods to treating hemoglobinopathies

• US 10,287,588 B2
• Filed: 04/24/2015
• Issued: 05/14/2019
• Est. Priority Date: 04/25/2014
• Status: Active Grant

• Current Assignee: Children's Medical Center Corporation
• Original Assignee: Children's Medical Center Corporation
• Inventors: Milsom, Michael, Williams, David A., Gregory, Richard
• Primary Examiner(s): Chong, Kimberly
• Application Number:
  US15/306,527
• Publication Number:
  US 20170218372A1
• Time in Patent Office: 1,481 Days
• Field of Search: None
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/1135 against oncogenes or tumor suppressor genes
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/141 MicroRNAs, miRNAs
• CPC Class Codes: C12N 2310/3519 Fusion with another nucleic acid
• CPC Class Codes: C12N 2310/531 Stem-loop; Hairpin
• CPC Class Codes: C12N 2740/10032 Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
• CPC Class Codes: C12N 2740/10043 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

Abstract

Embodiment herein provide specially designed synthetic BCL11A-targeting microRNAs for RNA polymerase II expression, and methods of use to treat hemoglobinopathies such as sickle cell disease or thalassemia by increasing the expression levels of fetal hemoglobin levels. In particular illustrative embodiment, the present invention provides, in part, improved compositions and methods for achieving gene therapy in hematopoietic cells and hematopoietic precursor cells, including erythrocytes, erythroid progenitors, and embryonic stem cells. The invention further provides improved gene therapy methods for treating hematopoietic-related disorders.

Claims

1. A synthetic BCL11A microRNA comprising:

a) a first BCL11A segment, a loop segment; and

b) a second BCL11A segment arranged in tandem in a 5′

to 3′

direction, wherein the loop segment is between and directly linked to the first and second BCL11A segments,

wherein the first BCL11A segment starts with a -GCGC- at the 5′

end and the second BCL11A segment ends with a -GCGC- at the 3′

end, and

wherein the first BCL11A segment is complementary to the second BCL11A segment so that the first and second BCL11A segments base pair to form a hairpin loop with the loop segment forming the loop portion of the hairpin loop thus formed.

2. The synthetic BCL11A microRNA of claim 1, wherein the first and second BCL11A segments are about 18 to 25 nucleotides long.

3. The synthetic BCL11A microRNA of claim 1, wherein the first BCL11A segment contains a sequence derived from a BCL11A mRNA sequence.

4. The synthetic BCL11A microRNA of claim 1, wherein the first BCL11A segment comprises the nucleotide sequence set forth in any one of SEQ ID NOs:

57, 58, 59, 60, 61, 62, 63, 64, and 65.

5. The synthetic BCL11A microRNA of claim 1, wherein the first BCL11A segment comprises the nucleotide sequence set forth in SEQ ID NO:

61.

6. The synthetic BCL11A microRNA of claim 1, wherein the loop segment is derived from a microRNA.

7. The synthetic BCL11A microRNA of claim 1, wherein the loop segment is derived from a microRNA selected from the group consisting of:

miR-142, miR-155, miR-181 and miR-223.

8. The synthetic BCL11A microRNA of claim 1, wherein the loop segment is derived from a microRNA selected from miR-223.

9. The synthetic BCL11A microRNA of claim 1, wherein the loop segment comprises the polynucleotide sequence set forth in SEQ ID NO:

68.

10. The synthetic BCL11A microRNA of claim 1, wherein the microRNA comprises a polynucleotide sequence set forth in any one of SEQ ID NOs:

34, 35, 36, 37, 38, 39, 40, 41, and 42.

11. The synthetic BCL11A microRNA of claim 1, wherein the microRNA comprises a polynucleotide sequence set forth in SEQ ID NO:

39.

12. A composition comprising the synthetic BCL11A microRNA of any one of claims 1-3 and 4-10.

13. A pharmaceutical composition comprising the synthetic BCL11A microRNA of any one of claims 1-3 and 4-10 and a pharmaceutically acceptable carrier or diluent.