# Primary Cilia-Dependent Mechanisms of Lymphangiogenesis

> **NIH NIH P20** · SOUTH DAKOTA STATE UNIVERSITY · 2022 · $351,335

## Abstract

PROJECT SUMMARY
Properly patterned lymphatic vessels are critical regulators of fluid and cellular traffic to support an effective
inflammation and wound healing response. However, the mechanisms that govern lymphatic vessel patterning
are not fully understood, and new therapeutics are needed to treat dysregulations of lymphangiogenesis, for
example in neovascularized corneal transplant rejection, lymphedema, and tumor-associated
lymphangiogenesis. The long-term goal of this research program is to develop lymphatic-targeted strategies to
promote wound healing and prevent or reverse refractory inflammation. In the course of work examining
lymphatic vessel pathology, the current investigators recently identified primary cilia on lymphatic endothelial
cells in vitro and in vivo. In other cell types, this organelle is a hub of cell signaling and regulates cellular
processes such as proliferation, cell polarity, and migration. Preliminary data in knockout mouse models of
IFT20, an intraflagellar transport protein important for primary cilia function, show that loss of primary cilia
exacerbates lymphangiogenesis. This suggests that previously unappreciated primary cilia-dependent
mechanisms may complement and modulate canonical mechanisms of lymphangiogenesis regulation. The
central hypothesis of this proposal is that unknown primary cilia signaling pathways negatively regulate
excessive lymphangiogenesis by suppressing sprouting, proliferation, and migration to form vasculature
appropriately tuned to local demand. This hypothesis will be tested with two specific aims: (1) Identify the
cellular mechanisms by which loss of primary cilia dysregulates lymphangiogenesis; and (2) Identify lymphatic
endothelial cell signaling pathways that are disrupted by loss of primary cilia. In aim 1, intravital imaging in
mice with IFT20 knockout and tdTomato reporter fluorescence along with in vitro lymphangiogenesis assays
will be used to identify the timing and extent of dysregulation of specific lymphatic endothelial cell activities that
comprise a functional lymphangiogenesis response, such as sprouting, cell-cell junction organization, and
chemotaxis. In aim 2, two next generation sequencing technologies, RNA-Seq and spatial transcriptomics, will
be used in combination with pathway analysis and gene ontology strategies and rational hypothesis-driven
prioritization of hits to identify candidate primary cilia-dependent regulators of inflammation-associated corneal
lymphangiogenesis. Candidates will be validated in vitro in CRISPR knockout lymphatic endothelial cell lines.
The combination of intravital imaging, transcriptomics, gene editing, and a panel of in vitro lymphangiogenesis
assays is an innovative strategy to comprehensively interrogate the cellular mechanistic basis of primary cilia
regulation of lymphangiogenesis. The proposed project is significant because it will provide the first
mechanistic insight into primary cilia function in lymphatic endothelial c...

## Key facts

- **NIH application ID:** 10335542
- **Project number:** 1P20GM135008-01A1
- **Recipient organization:** SOUTH DAKOTA STATE UNIVERSITY
- **Principal Investigator:** Darci M. Fink
- **Activity code:** P20 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $351,335
- **Award type:** 1
- **Project period:** 2022-03-20 → 2027-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10335542

## Citation

> US National Institutes of Health, RePORTER application 10335542, Primary Cilia-Dependent Mechanisms of Lymphangiogenesis (1P20GM135008-01A1). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10335542. Licensed CC0.

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