Improvements in Oligodendroglial Cell Culturing Methods and in Methods for Treating Neurodegenerative Disorders by Using Thyroid Hormones or Analogues

• US 20170342380A1
• Filed: 12/24/2015
• Published: 11/30/2017
• Est. Priority Date: 12/24/2014
• Status: Active Grant

• Current Assignee: Neuorphan Pty, Ltd.
• Original Assignee: Neuorphan Pty, Ltd.
• Inventors: Petratos, Steven, Azari, Michael Farzad, Lee, Jae Young, Kim, Min Young
• Granted Patent: US 10,640,748 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 35/545 Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
• CPC Class Codes: C12N 2501/395 Thyroid hormones
• CPC Class Codes: C12N 2502/081 neurons
• CPC Class Codes: C12N 2502/086 glial cells
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0618 Cells of the nervous system
• CPC Class Codes: C12N 5/0622 Glial cells, e.g. astrocytes, oligodendrocytes; Schwann cells

Abstract

The present invention relates to methods of treating or ameliorating certain neurodegenerative disorders (namely, dysmyelinating and demyelinating disorders) in patients in need of such treatment or amelioration. The invention provides methods of treating or ameliorating a patient in need of such treatment and includes the administration to the patient of: (a) thyroid hormones or thyroid hormone analogues; (b) cell replacement therapies involving the use of homogenous Oligodendrocyte Precursor Cells derived from embryonic stem cells that have been treated with thyroid hormones or thyroid hormone analogues; (c) gene therapy to correct mutated genes in vivo; or (d) a combination of two or more of (a), (b) and (c). The invention also provides compositions and formulations of thyroid hormones and thyroid hormone analogues for use in treating or ameliorating such disorders.

Claims

1-36. -36. (canceled)

37. A method of differentiating a vertebrate embryonic stem cell into a vertebrate oligodendrocyte precursor cell, the method comprising culturing the vertebrate embryonic stem cell in a culture medium comprising 3,5-diiodothyropropionic acid (DITPA) at a concentration which is effective to cause the vertebrate embryonic stem cell to differentiate into an oligodendrocyte precursor cell.

38. The method according to claim 37, wherein the vertebrate embryonic stem cell is mammalian.

39. The method according to claim 37, wherein the vertebrate embryonic stem cell is human.

40. The method according to claim 37 wherein the vertebrate embryonic stem cell is an hESC-derived Nkx2.1-positive reporter cell or a Hes3 reporter cell.

41. The method according to claim 37 wherein the vertebrate embryonic stem cell is an H9 human embryonic stem cell.

42. The method according to claim 37, wherein the vertebrate oligodendrocyte precursor cell is capable of myelinating a neuron in a human subject.

43. The method according to claim 37, comprises culturing the vertebrate embryonic stem cell in the culture medium comprising DITPA for up to 3 months.

44. The method according to claim 37, wherein the culture medium comprises between I ng/mL and 100 ng/mL DITPA.

45. The method according to claim 37, further comprising:

adding FGF2 to the culture medium and culturing so that a three-dimensional mass of cells forms;

adding FGF2, FGF4 and noggin to the culture medium and culturing so that an embryoid body forms from the three-dimensional mass;

adding sonic hedgehog (shh), and optionally FGF2, to the culture medium and culturing so that neural precursor cells form from cells of the embryoid body;

adding EGF to the culture medium and culturing so that glial precursor cells form from the neural precursor cells;

adding PDGF to the culture medium and culturing so that oligodendrocyte precursor cells form glial precursor cells; and

culturing the oligodendrocyte precursor cells in the culture medium to form post-mitotic cells capable of myelination.

46. A method as claimed in claim 37, further comprising:

adding EGF to the culture medium and culturing so that glial precursor cells form;

adding PDGF to the culture medium and culturing so that oligodendrocyte precursor cells form; and

culturing the oligodendrocyte precursor cells in the culture medium to form post-mitotic cells capable of myelination,

wherein the vertebrate embryonic stem cell is an hESC-derived Nkx2.1-positive reporter cell or a Hes3 reporter cell.

50. A vertebrate oligodendrocyte precursor cell differentiated according to a method of claim 37.

51. A cell line comprising a vertebrate oligodendrocyte precursor cell differentiated according to a method of claim 37.

47. A method of treating or ameliorating a neurological disorder associated with slc16A2 gene or MCT8 deficiency or impairment in a patient in need of such treatment or amelioration, the method comprising administering to the patient an effective amount of a vertebrate oligodendrocyte precursor cell differentiated from a vertebrate embryonic stem cell by a method comprising culturing the vertebrate embryonic stem cell in a culture medium comprising a thyroid hormone or a thyroid hormone analogue at a concentration which is effective to cause the vertebrate embryonic stem cell to differentiate into an oligodendrocyte precursor cell.

48. The method according to claim 47, wherein the thyroid hormone or thyroid hormone analogue is DITPA.

49. The method according to claim 47, wherein the thyroid hormone or thyroid hormone analogue is DITPA and the culture medium comprises between I ng/mL and 100 ng/mL DITPA.