# MicroBRAINS: Bioengineered Human Neural Circuits for Aging Research

> **NIH NIH R21** · METHODIST HOSPITAL RESEARCH INSTITUTE · 2020 · $201,875

## Abstract

PROJECT SUMMARY-ABSTRACT
 Our goal is to generate and validate an innovative 3D cellular model of the nervous system from human
pluripotent stem cells as a new tool for aging research. The current state of the art for in vitro modeling is
insufficient because multicellular human neural circuits in culture conditions remain in immature states for
prolonged periods of time. To overcome this barrier, we are proposing to produce a Microassembly of
Bioengineered, Rapid, All-Inducible Neural System (µBRAINS). We will utilize our recently optimized protocols
to produce consistent 3D cocultures generated from novel transgenic cell lines with transcription activator-like
effector nuclease technology that we have generated in preliminary data. In Aim 1 neurons, astrocytes and
oligodendrocytes will be individually produced from human pluripotent stem cells by induction of neurogenin 2,
Sox9 in combination with NFIA, and Sox10, respectively. We will assemble these cell types together in defined
3D coculture spheres and assess neural circuit maturation over time. Maturation will be determined by increased
presence of myelinated fibers and onset of synchronous network activity. Activity will be measured by
multielectrode array recordings as well as live calcium indicator imaging. These studies will determine the optimal
conditions for producing the functional µBRAINS. In Aim 2, we will test whether regulated neuronal activity will
enforce maturation in a rapid and consistent manner. We will utilize a novel optogenetic cell line generated in
preliminary studies to determine the effect of light-mediated stimulation of neurons upon circuit maturation within
the µBRAINS. The results from these studies will reveal if evoked neuronal activity accelerates production of
mature circuits. In Aim 3, we will test whether forced expression of the transcription factor NFIX in astrocytes will
induce aging phenotypes by performing assays for cellular aging and by measuring the effect upon myelination
and neural network activity of cocultured cells. Altogether, we expect to deliver novel cell lines and an optimized
protocol to systematically generate a human neural circuit model system with aging phenotypes. The µBRAINS
can be a breakthrough tool for use throughout the research community for disease modeling and screening drug
interventions in the context of age-related conditions such as Alzheimer’s disease.

## Key facts

- **NIH application ID:** 9985696
- **Project number:** 5R21AG064567-02
- **Recipient organization:** METHODIST HOSPITAL RESEARCH INSTITUTE
- **Principal Investigator:** Robert Conrad Krencik
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $201,875
- **Award type:** 5
- **Project period:** 2019-07-01 → 2022-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9985696

## Citation

> US National Institutes of Health, RePORTER application 9985696, MicroBRAINS: Bioengineered Human Neural Circuits for Aging Research (5R21AG064567-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/9985696. Licensed CC0.

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