# Advanced conformational changes in NOTCH3 and cerebrovascular disease severity

> **NIH VA I01** · VETERANS HEALTH ADMINISTRATION · 2024 · —

## Abstract

ABSTRACT
Neurodegenerative disease is a major cause of death and permanent disability in Veterans;
vascular disease of the brain plays an outsized role in the progression of neurodegenerative
diseases including Alzheimer’s Disease and Related Dementias (ADRD). On its own, vascular
disease of the brain can cause one form of ADRD, vascular cognitive impairment and dementia
(VCID). In collaboration with disorders such as Alzheimer’s disease, brain vascular disease
significantly augments the pace and severity of dementing disorders. Unfortunately, effective
treatments that target vascular disease of the brain are currently not available. As such, there
is an unmet need to uncover the molecular mechanisms of brain vascular disease. The most
common inherited cause of brain vascular disease is cerebral autosomal dominant arteriopathy
with subcortical infarcts and dementia (CADASIL). CADASIL is an outstanding model for
understanding pathology of brain blood vessels and is caused by mutations in NOTCH3 that are
predicted to alter cysteine number. We have discovered a sequence of changes in the
configuration of NOTCH3 in association with CADASIL mutations. First, NOTCH3 in CADASIL
is more likely to harbor multiple reduced cysteines (a form called mrc-N3). Second, NOTCH3
that is multiply reduced undergoes cleavage at the N-terminus that releases a 41-amino acid
peptide, the NOTCH3 N-terminal fragment (NTF). Based on the molecular process driving NTF
generation, in new experiments, we describe the identification of a second cleavage event that
is predicted to result in a second 41-amino acid peptide NTF2. Antibodies generated to NTF2
demonstrate that CADASIL vessels are enriched in this cleavage product. Moreover, a novel
monoclonal antibody has been identified that recognizes a conformation of NTF2 that is only
expressed in a subset of vessels (NTF2*). In this proposal, we will test the hypothesis that
NTF2* is associated with advanced pathology of CADASIL blood vessels. We will also test if
NTF2* is deposited in a gradient from the surface to the deep white matter, which would point to
the possibility that NTF2* is a predictable marker for vascular disease progression.

## Key facts

- **NIH application ID:** 10726584
- **Project number:** 5I01BX003824-07
- **Recipient organization:** VETERANS HEALTH ADMINISTRATION
- **Principal Investigator:** Michael M Wang
- **Activity code:** I01 (R01, R21, SBIR, etc.)
- **Funding institute:** VA
- **Fiscal year:** 2024
- **Award amount:** —
- **Award type:** 5
- **Project period:** 2017-04-01 → 2029-09-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10726584

## Citation

> US National Institutes of Health, RePORTER application 10726584, Advanced conformational changes in NOTCH3 and cerebrovascular disease severity (5I01BX003824-07). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10726584. Licensed CC0.

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