US 12,169,196 B2
Human iPSC-based drug testing platform for muscular dystrophy
Gabsang Lee, Baltimore, MD (US); Kathryn Wagner, Baltimore, MD (US); and Congshan Sun, Baltimore, MD (US)
Assigned to The Johns Hopkins University, Baltimore, MD (US)
Filed by The Johns Hopkins University, Baltimore, MD (US)
Filed on Apr. 3, 2020, as Appl. No. 16/840,302.
Claims priority of provisional application 62/828,991, filed on Apr. 3, 2019.
Prior Publication US 2020/0355674 A1, Nov. 12, 2020
Int. Cl. G01N 33/50 (2006.01); C12N 5/074 (2010.01); C12N 5/077 (2010.01)
CPC G01N 33/5061 (2013.01) [C12N 5/0658 (2013.01); C12N 5/0696 (2013.01); C12N 2501/60 (2013.01); C12N 2501/602 (2013.01); C12N 2501/604 (2013.01); C12N 2501/606 (2013.01); C12N 2503/02 (2013.01); C12N 2506/1307 (2013.01); C12N 2506/45 (2013.01)] 13 Claims
 
1. A method of screening for candidate therapeutic agents, comprising: obtaining fibroblasts from a subject diagnosed as having a muscular dystrophy and generating induced pluripotent stem cells (iPSCs); culturing the iPSCs; differentiating the iPSCs to generate myoblasts; contacting the myoblasts with a candidate therapeutic agent; culturing the myoblasts with a detectably labeled anti-myosin heavy chain antibody; and, imaging and analyzing the myoblasts generated from the iPSCs of the subject diagnosed with muscular dystrophy as compared to myoblasts generated from a healthy subject's iPSCs, wherein the analysis comprises measuring average length of myoblasts and expression of myosin heavy chain (MyHC) polypeptides as compared to positive and negative controls; and wherein the average length of myoblasts is determined by: cell average length+0.3*MyHC (myosin heavy chain); thereby, screening for the candidate therapeutic agent.