Mapping the spatial and temporal responses of hESC-derived microglia to repeat mild closed head injury to identify therapeutic targets and mechanisms
Grant Award Details
Grant Type:
Grant Number:
DISC0-14447
Investigator(s):
Disease Focus:
Human Stem Cell Use:
Award Value:
$1,339,685
Status:
Active
Grant Application Details
Application Title:
Mapping the spatial and temporal responses of hESC-derived microglia to repeat mild closed head injury to identify therapeutic targets and mechanisms
Public Abstract:
Research Objective
We will generate an RNA activation map of human stem cell derived microglia activation states following brain injury to then test a new gene-edited microglia peptide delivery mechanism.
Impact
Bottlenecks with the time and sex-dependent human microglia responses to repeat mild closed head injury and questions surrounding the delivery and efficacy of a trophic factor as a therapeutic.
Major Proposed Activities
We will generate an RNA activation map of human stem cell derived microglia activation states following brain injury to then test a new gene-edited microglia peptide delivery mechanism.
Impact
Bottlenecks with the time and sex-dependent human microglia responses to repeat mild closed head injury and questions surrounding the delivery and efficacy of a trophic factor as a therapeutic.
Major Proposed Activities
- Generate an RNA map of the human microglial response to repeat mild concussions at multiple post-injury timepoints on mice transplanted with microglia derived from male and female stem cells.
- Use microscopy to histologically validate the microglia populations identified in Activity 1 express candidate genes from the early vs late and/or male vs female injury responses.
- CRISPR edit male and female stem cell lines to append the sequence for a secreted trophic factor coupled to a reporter peptide at the end of the selected injury-response genes found in Activity 1.
- Verify that the microglia generated from CRISPR editing are different than the parental lines as designed in Activity 3 (quality control).
- Stimulate the CRISPR edited microglial lines in cell culture to confirm that the trophic factor/reporter peptide responds to injury and is secreted.
- Perform repeat mild concussions on mice transplanted with the CRISPR engineered microglial cell lines and verify that the inserted trophic factor/reporter peptide is delivered to the injury site.
Statement of Benefit to California:
This research could benefit California by discovering genes that are specifically upregulated in microglia in response to brain injury. Editing a gene to deliver a trophic factor to the injury could be a pathway for treating concussion/closed head injuries or even more severe traumatic brain injuries (which effect >230,000 Californians/year and cost CA ~9.6 billion every year). This bottleneck could also open the door for stem cell based treatments of a variety of neurological conditions.