# High throughput assaying of circuit activity and connectivity in brain organoids

> **NIH NIH RF1** · HARVARD UNIVERSITY · 2020 · $2,509,021

## Abstract

Experimental models of the human developing brain are needed to investigate human-specific aspects of
brain development, evolution, and neurological disease. Progress in the field has been hampered by the lack of
models, considering that the endogenous developing human brain cannot be directly investigated; animal models
often fail to recapitulate human disorders and cannot feasibly be used to study complex polygenic states
spanning many genes. While reductionist in nature, stem-cell derived 3D human brain organoids offer a first-of-
its-kind opportunity to study processes of human brain formation and wiring that are otherwise not accessible.
However, there is an unmet need for organoid models that are cellularly complete and reproducible and for
methodology to decode the establishment, connectivity and dynamics of neural circuits in organoids, at scale
and with high fidelity. If we could map the activity and connectivity of organoids at scale, both to
understand circuit function/dysfunction and to guide further development of organoids, we could close
the loop on organoid design and application. Towards this goal, we have developed many molecular and
imaging tools for high-throughput analysis of neural activity and connectivity, which we propose to apply to new,
next-generation organoid models. Here, we propose a collaborative approach among four groups (Arlotta - brain
organoids and human brain development; Boyden - circuit physiology and neural imaging technology; Lewis -
material science and bioengineering and Insoo Hyun- bioethics) to pioneer a robust organoid system that
combines the development of vascularized brain organoids incorporating more complete cell diversity and
maturation with advanced high-throughput functional molecular and imaging tools to enable interrogation of
circuit activity, connectivity, and molecular changes in cells participating in physiologically relevant circuits. We
will build on a highly reproducible brain organoid model that we recently developed to promote the generation of
cell types that are currently absent in organoids but needed for circuit maturation, refinement, and functionality.
This work is intended to generate more advanced organoid models designed to promote maturation and robust
network activity. In parallel, we will develop a pipeline to record neural activity from intact organoids using all-
optical-electrophysiology techniques at scale, and optimize epitope-based barcoding and expansion microscopy
to enable molecularly-annotated connectomics of brain organoids. The work proposed here will enable the use
of human organoid models to study human circuit formation, plasticity, and function, analyses that are currently
hampered by the lack of technologies and assays for high-throughput measurements of circuit physiology and
connectivity in organoids. Beyond the work proposed here, these methods will directly enable investigation into
how disease states alter information processing in the brain; for ...

## Key facts

- **NIH application ID:** 10049677
- **Project number:** 1RF1MH123977-01
- **Recipient organization:** HARVARD UNIVERSITY
- **Principal Investigator:** Paola Arlotta
- **Activity code:** RF1 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $2,509,021
- **Award type:** 1
- **Project period:** 2020-09-01 → 2024-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10049677

## Citation

> US National Institutes of Health, RePORTER application 10049677, High throughput assaying of circuit activity and connectivity in brain organoids (1RF1MH123977-01). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10049677. Licensed CC0.

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