# Generation of Neurons by Force-Mediated Epigenetic Mechanisms through Manipulation of Intrinsic Mechanoregulators

> **NIH NIH K01** · ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI · 2024 · $240,354

## Abstract

PROJECT SUMMARY
 There is a great interest in deriving induced neuronal (iN) cells from human pluripotent stem cells (hPSCs) for
central nervous system disease modeling and drug discovery. However, current differentiation methods are
laborious and inefficient, thus, identifying new ways to improve neuronal differentiation is desirable. Strikingly, in
my most recent publication I discovered that knockout of the axon guidance receptor Plexin-B2 in hNPCs
reduced cell stiffness, increased Yes-Associated Protein 1 (YAP) phosphorylation, and accelerated neuronal
differentiation. As a critical downstream mechanism, my preliminary data points to a pathway of Plexin-B2 altered
mechanoregulation affecting lamin-mediated nuclear envelope architecture and localization of repressive histone
marks at genomic loci containing neuron-specific genes.
 Based on these data, I will investigate an intrinsic ‘force-based’ method to generate iNs by manipulating
Plexin-B2, Angiomotin (AMOT; a mechanosensitive component of Hippo pathway), and YAP in both 2D and 3D
neuronal cultures and examine the link between Plexin-B2 and Hippo/YAP mechano-signaling in regulating
lamin-mediated nuclear physical properties, force transmission to the nuclear envelope, and changes in
chromatin accessibility during neural induction. To carry out this investigation, I propose the following aims: #1.
Investigate the impact of Plexin-B2-altered cell mechanics on neuronal differentiation, by optimizing the
differentiation method to generate iNs by both Plexin-B2 knockdown and knockout in multiple hPSC lines. I will
test the robustness, stability, and functionality of iNs by evaluating transcriptional identity, neuronal lineage,
maturation, and survival markers, electrophysiological properties, and neurotransmitter release. #2. Investigate
a mechanochemical loop between Plexin-B2 and YAP during neuronal differentiation, by performing a
comprehensive gain- and loss-of-function epistasis studies to map out the relationships between Plexin-B2, YAP,
and AMOT during neural induction and evaluate their impact on enhancing neuronal differentiation, maturation,
survival, electrophysiological functionality, and expression of major neuronal lineage markers. #3. Investigate
lamin-mediated nuclear architecture and chromatin interactions in Plexin-YAP mechano-regulation of
neuronal differentiation, by performing a comprehensive gain- and loss-of-function epistasis studies to validate
lamin as the mediator of a Plexin-B2 and Hippo/YAP intrinsically-regulated mechano-axis to promote neuronal
differentiation. I will then evaluate the role of Plexin-B2 / Hippo/YAP / lamins in releasing genomic loci harboring
neuron-specific genes from repressive chromatin at nuclear periphery to interior regions with open chromatin.
 This K01 Award will provide me training in neuronal electrophysiology, transcriptomics and epigenomics,
lab management and mentoring skills, as well as time and mentoring to develop and write my fir...

## Key facts

- **NIH application ID:** 10818502
- **Project number:** 5K01NS127948-02
- **Recipient organization:** ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
- **Principal Investigator:** Chrystian Junqueira Alves
- **Activity code:** K01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $240,354
- **Award type:** 5
- **Project period:** 2023-04-01 → 2028-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10818502

## Citation

> US National Institutes of Health, RePORTER application 10818502, Generation of Neurons by Force-Mediated Epigenetic Mechanisms through Manipulation of Intrinsic Mechanoregulators (5K01NS127948-02). Retrieved via AI Analytics 2026-05-28 from https://api.ai-analytics.org/grant/nih/10818502. Licensed CC0.

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