US 11,674,952 B2
Embryonic cell-based therapeutic candidate screening systems, models for Huntington's Disease and uses thereof
Ali Brivanlou, NY, NY (US); Albert Ruzo, New York, NY (US); Alessia Deglincerti, New York, NY (US); Tomomi Haremaki, New York, NY (US); and Fred Etoc, New York, NY (US)
Assigned to The Rockefeller University, New York, NY (US)
Appl. No. 16/79,755
Filed by The Rockefeller University, New York, NY (US)
PCT Filed Feb. 24, 2017, PCT No. PCT/US2017/019529
§ 371(c)(1), (2) Date Aug. 24, 2018,
PCT Pub. No. WO2017/147536, PCT Pub. Date Aug. 31, 2017.
Claims priority of provisional application 62/300,056, filed on Feb. 25, 2016.
Claims priority of provisional application 62/299,544, filed on Feb. 24, 2016.
Prior Publication US 2019/0195863 A1, Jun. 27, 2019
Int. Cl. C12Q 1/02 (2006.01); G01N 33/50 (2006.01); C12Q 1/18 (2006.01); C12N 15/09 (2006.01); C12N 5/0735 (2010.01); A61K 31/506 (2006.01); A61K 45/06 (2006.01); C07K 14/47 (2006.01)
CPC G01N 33/5073 (2013.01) [A61K 31/506 (2013.01); A61K 45/06 (2013.01); C07K 14/47 (2013.01); C12N 5/0606 (2013.01); C12N 15/09 (2013.01); C12Q 1/02 (2013.01); C12Q 1/025 (2013.01); C12Q 1/18 (2013.01); C12N 2503/02 (2013.01); C12N 2510/00 (2013.01)] 17 Claims
 
1. A method for identifying therapeutic candidates for treatment of Huntington's Disease (HD) comprising:
(a) contacting a test substance with each of:
(i) a first clonal population of human embryonic stem cells comprising a wild type Huntingtin (HTT) gene, and
(ii) a second clonal population of human embryonic stem cells isogenic to the first population, wherein the cells in the second population are genetically modified to comprise at the endogenous HTT gene locus a nucleotide segment, wherein the nucleotide segment, upon expression, results in a polyQ repeat peptide segment comprising at least 36 or at least 40 glutamine residues in the N-terminal region of the HTT protein,
wherein the first clonal population of human embryonic stem cells and the second clonal population of embryonic stems cells are each cultured on a micropatterned confined surface pretreated with an adherent substrate; and
(b) determining whether the test substance:
(i) wholly or partially reverses a perturbed organization of the cells of the second clonal population or cells differentiated therefrom; and
(ii) reverts a change in three-dimensional cellular geometry associated with a micropattern signature in the second clonal population or cells differentiated therefrom.