Aiolos, Helios, Daedalos and Ikaros: genes, polypeptides, regulatory elements and uses thereof

• US 20050130265A1
• Filed: 07/06/2004
• Published: 06/16/2005
• Est. Priority Date: 09/14/1992
• Status: Active Grant

• Current Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Original Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Inventors: Morgan, Bruce A., Georgopoulos, Katia, Kelley, Clair
• Granted Patent: US 7,196,170 B2
• Time in Patent Office: Days
• US Class Current: 435/69.100
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/03 Animal model, e.g. for test or diseases
• CPC Class Codes: A01K 2267/0381 Animal model for diseases of the hematopoietic system
• CPC Class Codes: A01K 67/0276 Knock-out vertebrates
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07K 14/4705 stimulating, promoting or activating activity
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2800/30 Vector systems comprising sequences for excision in presence of a recombinase, e.g. loxP or FRT
• CPC Class Codes: C12N 2830/008 cell type or tissue specific enhancer/promoter combination
• CPC Class Codes: G01N 2333/4706 stimulating, promoting or activating activity
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism
• CPC Class Codes: G01N 33/56966 Animal cells
• CPC Class Codes: G01N 33/56972 White blood cells
• CPC Class Codes: G01N 33/581 with enzyme label (including co-enzymes, co-factors, enzyme inhibitors or substrates)
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere

Abstract

Provided herein are (a) Aiolos gene, Aiolos polypeptides, Aiolos homodimers, Aiolos/Ikaros heterodimers and methods of using Aiolos nucleic acids and polypeptides; (b) Helios gene, Helios polypeptides, Helios homodimers, Helios/Ikaros heterodimers, Helios/Aiolos heterodimers, and methods of using Helios nucleic acids and polypeptides; (c) Daedalos nucleic acids, Daedalos polypeptides, and other related molecules and methods of making and using the same; and (d) Ikaros regulatory elements and uses thereof.

Claims

1. A substantially pure polypeptide which is at least 60% homologous to a Helios polypeptide.

2. The Helios polypeptide of claim 1 comprising the amino acid sequence of SEQ ID NO:

24, 26, or 28.

3. A fragment of the Helios polypeptide of claim 1 which is at least 50 amino acids in length.

4. The pure preparation of claim 1, wherein the Helios polypeptide has the following properties:

(a) it can form a dimer with an Helios, Aiolos, or Ikaros polypeptide;

(b) it is expressed in hematopoietic stem cells;

(c) it has a molecular weight of approximately 64 kDa or 68 KDa;

(d) it has at least one zinc finger domain; and

(e) it is a transcriptional activator of a lymphoid gene.

6. A method of making a Helios polypeptide, having at least one biological activity of a naturally occurring Helios polypeptide including altering the sequence of one or more residues of the polypeptide of claim 1, and testing the altered polypeptide for the desired activity.

7. A method for treating an animal for a disorder comprising administering a therapeutically-effective amount of a Helios polypeptide of claim 6.

5. A purified preparation of an anti-Helios antibody.

8. An isolated Ikaros transcriptional control region comprising one or more Ikaros regulatory element found in a lymphoid-specific DNaseI HSS cluster selected from:

a 9 kb BamHI fragment of a lymphoid-specific DNaseI hypersensitive site (HSS) of the mouse or human Ikaros locus (α

cluster);

a 5.9 kb BamHI/EcoRI fragment of a lymphoid-specific DNaseI HSS of the mouse or human Ikaros locus (β

cluster);

a 5 kb EcoRI fragment of a lymphoid-specific DNaseI HSS of the mouse or human Ikaros locus (γ

cluster);

a 4.2 kb EcoRI fragment of a lymphoid-specific DNaseI HSS of the mouse or human Ikaros locus (δ

cluster);

a 11 kb BamHI fragment of a lymphoid-specific DNaseI HSS of the mouse or human Ikaros locus (ε

cluster);

a 13.5 kb EcoRI fragment of a lymphoid-specific DNaseI HSS of the mouse or human Ikaros locus (ζ

cluster);

a 3.7 kb XbaI fragment of a lymphoid-specific DNaseI HSS of the mouse or human Ikaros locus (η

cluster); and

7.5 kb BamHI fragment of a lymphoid-specific DNaseI HSS of the mouse or human Ikaros locus (θ

cluster).

9. A construct comprising an Ikaros transcriptional control region of claim 8 operably linked to a sequence encoding a reporter molecule.

10. The DNA construct of claim 9, wherein the reporter molecule is a reporter molecule which can luminesce or fluoresce.

11. A method for treating an animal for a disorder comprising administering a therapeutically-effective amount of an Aiolos polypeptide, a cell selected for the expression of a product of the Aiolos gene, or a nucleic acid encoding an Aiolos peptide to the animal.

12. The method of claim 11 wherein the Aiolos polypeptide has the following properties:

(a) it can form a dimer with an Aiolos or Ikaros polypeptide;

(b) it is expressed in committed lymphoid progenitors;

(c) it is expressed in committed T and B cells;

(d) it has a molecular weight of approximately 58 kD;

(e) it has at least one zinc finger domain;

(f) it is not expressed in stem cells; and

(g) it is a transcriptional activator of a lymphoid gene.

13. The method of claim 11 wherein the Aiolos polypeptide has at least one biological activity of a naturally occurring Aiolos polypeptide, and the polypeptide has the sequence of SEQ ID NO:

8 with one or more altered amino acids.

14. A method of treating a neural cell related disorder in a subject, comprising:

providing a subject having a neural cell related disorder; and

modulating expression, levels or activity of Daedalos in a cell of the subject, to thereby treat the disorder.

15. The method of claim 14, wherein expression, levels or activity of Daedalos is inhibited.

16. The method of claim 15, wherein the expression, levels or activity of Daedalos is inhibited by administering to the subject an agent selected from the group consisting of:

a Daedalos binding protein that inhibits a Daedalos activity;

an antibody to Daedalos that inhibits a Daedalos activity;

a mutated Daedalos or fragment thereof that inhibits a Daedalos activity;

a Daedalos nucleic acid molecule that inhibits expression of Daedalos; and

a small molecule that inhibits transcription or activity of Daedalos.

17. The method of claim 14, wherein the disorder is cancer.

18. The method of claim 14, wherein expression, levels or activity of Daedalos is increased.