|
| Status |
Public on Nov 11, 2021 |
| Title |
Perturb-Seq using T2D islets |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
Type 2 diabetes is associated with defective insulin secretion and reduced β-cell mass. Available treatments provide a temporary reprieve, but secondary failure rates are high, making insulin supplementation necessary. Reversibility of b-cell failure is a key translational question. Here, we reverse-engineered and interrogated pancreatic islet-specific regulatory networks to discover T2D-specific subpopulations characterized by metabolic-inflexibility and endocrine-progenitor/stem cell features. Single-cell gain- and loss-of-function and glucose-induced Ca++ flux analyses of top candidate MR in islet cells validated transcription factor BACH2 and associated epigenetic effectors as a key driver of T2D cell states. BACH2 knockout in T2D islets reversed cellular features of the disease, restoring a non-diabetic phenotype. BACH2-immunoreactive islet cells increased ~4-fold in diabetic patients, confirming the algorithmic prediction of clinically relevant subpopulations. Treatment with a BACH inhibitor lowered glycemia and increased plasma insulin levels in diabetic mice, and restored insulin secretion in diabetic mice and human islets. The findings suggest that T2D-specific populations of failing b-cells can be reversed and indicate pathways for pharmacological intervention, including via BACH2 inhibition.
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| Overall design |
Adenovirus encoding CAS9_gRNAs targetting candidate TF with its corresponding DNA guide-barcode was infected in human T2D islets which are subjected to scRNA-seq.
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| Contributor(s) |
Son J, Ding H, Califano A, Accili D |
| Citation(s) |
34907913 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R01 DK064819 |
Mechanisms of Beta Cell Failure |
Trustees of Columbia University in the City of New York |
DOMENICO ACCILI |
| R35 CA197745 |
Elucidating the dependencies of tumor initiating and drug-resistant niches in human malignancies by genome- wide molecualr profiling of single cells |
Trustees of Columbia University in the City of New York |
ANDREA CALIFANO |
| P30 DK063608 |
Columbia Diabetes Research Center |
Trustees of Columbia University in the City of New York |
DOMENICO ACCILI |
| F32 DK117574 |
Analysis of beta cell dedifferentiation using scRNA-seq of human T2D |
Trustees of Columbia University in the City of New York |
Jinsook Son |
|
| Submission date |
Sep 19, 2019 |
| Last update date |
Oct 07, 2022 |
| Contact name |
Hongxu Ding |
| E-mail(s) |
hongxuding@arizona.edu
|
| Organization name |
University of Arizona
|
| Street address |
The University of Arizona
|
| City |
Tucson |
| State/province |
AZ |
| ZIP/Postal code |
85721 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
|
| Samples (4)
|
| GSM4087037 |
Perturb-Seq T2D donor (T2D7) ctrl Dataset 1 |
| GSM4087038 |
Perturb-Seq T2D donor (T2D7) ctrl Dataset 2 |
| GSM4087039 |
Perturb-Seq T2D donor (T2D7) Dataset 1 |
|
| Relations |
| BioProject |
PRJNA568200 |
| SRA |
SRP222610 |
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