Embryonic cell-based therapeutic candidate screening systems, models for huntington's disease and uses thereof

Abstract

Compositions and methods disclosed concern an isogenic population of in vitro human embryonic stem cells comprising a disease form of the Huntingtin gene (HTT) at the endogenous HTT gene locus in thegenome of the cell; wherein the disease form of the HTT gene comprises a polyQ repeat of at least 40 glutamines at the N-terminus of the Huntingtin protein (HTT). The cell lines of the disclosure comprise genetically-defined alterations made in the endogenous HTT gene that recapitulate Huntington's Disease in humans. Furthermore, the cell lines have isogenic controls that share a similar genetic background. Differentiating cell lines committed to a neuronal fate and fully differentiated cell lines are also provided and they also display phenotypic abnormalities associated with the length of the polyQ repeat of the HTT gene. These cell lines are used as screening tools in drug discovery and development to identify substances that fully or partially revert these phenotype abnormalities.

Description

[0001] background of the invention [0002] government rights [0003] The invention disclosed herein was made in part with government support under Grant Nos R01 GM101653 and R01 HD080699 awarded by the National Institutes of Health. The US Government has certain rights in this invention. [0004] related application [0005] This international patent application claims priority to U.S. Provisional Patent Application No. 62 / 299,544, filed February 24, 2016, and U.S. Provisional Patent Application No. 62 / 300,056, filed February 25, 2016. The entire disclosure of each of the above provisional patent applications is incorporated by reference. [0006] sequence listing [0007] This patent application contains a Sequence Listing filed electronically in ASCII format, and the Sequence Listing is hereby incorporated by reference in its entirety. Said ASCII copy, created on February 24, 2017, is named 11012-005430-WO0_SL.TXT and is 43,621 bytes in size. technical field [0008...

AI Technical Summary

Problems solved by technology

However, unlike the highly inbred mouse, the human system displays heterogeneity in the genetic background, making comparative studies difficult to interpret and generalize from individual cases

However, human iPSCs have been used to model human diseases, and although hiPSCs are a general model, their major drawbacks include: incomplete reprogramming, genomic instability, and different differentiation potentials depending on the epigenetic memory from which they originate

In addition, since these cells are derived from patients with mutations throughout their lifetime, there may be intrinsic alterations that impair cellular homeostasis

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