# Complement-Induced Endothelial Cell Activation by a Novel Rab5-ZFYVE21-SMURF2 Signaling Axis

> **NIH NIH R01** · YALE UNIVERSITY · 2021 · $82,038

## Abstract

PROJECT SUMMARY
 Transplant arteriosclerosis (TA) is a vaso-occlusive condition characterized by the formation of stenotic
lesions in arterial and microvascular beds of solid organ allografts that cause ischemic graft loss. Complement
(C’) are immune proteins involved in host defense that become pathologically activated on endothelial cells
(ECs) during TA. In TA, complement becomes activated by recipient-derived alloantibodies binding to donor
ECs. Upon activation complement proteins self-assemble to form membrane attack complexes (MAC),
heterodimers that insert into target EC surfaces as transmembranous pores.
 The pathologic effects of complement activation including assembly on ECs is well established,
however the precise molecular basis underlying these effects remains unclear, largely due to the lack of
relevant experimental models for studying MAC. To obviate this, we developed models for antibody-mediated
C’ activation on ECs in vitro and in vivo. Our protocols incorporated human biospecimens in order to
recapitulate TA phenotypes in patients. Using these protocols, we defined a novel signaling mechanism
causing EC activation and EC-mediated immune responses promoting TA.
 We have since built a collaborative infrastructure to obtain myriad patient biospecimens to support our
work. These patient specimens include ‘high’ panel reactive antibody sera, fresh human coronary arteries, sera
and PBMCs from prospectively enrolled patients, healthy PBMCs, TA biopsies, and biopsies from patients with
various complement-mediated conditions involving vascular inflammation. Collection of these specimens
involved collaborative interactions with the Tissue Typing Lab, Dept of Cardiothoracic Surgery, Dept of
Transplant Surgery, Apheresis Service, and various Depts of Pathology. Due to the broad and extensive use of
the materials above, my laboratory is exquisitely dependent on their ongoing collection for productivity.
 In this supplement, we propose a strategy to offset potential losses of productivity due to a critical life
event, i.e., the diagnosis of metastatic pancreatic adenocarcinoma in my father. Supplemental funding will be
used to hire a research technician who will possess the requisite skillsets and availability on nights and
weekends to collect patient specimens. A plan for hiring, training, evaluating, and mentoring this individual is
provided along with descriptions of current lab productivity, the critical life event, and a planned return to full
productivity at the end of the supplemental period. Use of supplemental funding will occur contemporaneously
with various mechanisms intended to maintain productivity both during the supplemental period and include
institutional commitments for FMLA, benefits, protected time, colleague support, and wellness resources.

## Key facts

- **NIH application ID:** 10294356
- **Project number:** 3R01HL141137-04S2
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** Dan Jane-Wit
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $82,038
- **Award type:** 3
- **Project period:** 2018-04-01 → 2023-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10294356

## Citation

> US National Institutes of Health, RePORTER application 10294356, Complement-Induced Endothelial Cell Activation by a Novel Rab5-ZFYVE21-SMURF2 Signaling Axis (3R01HL141137-04S2). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10294356. Licensed CC0.

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