# Defining the molecular mediators of cell morphogenesis in Trypanosoma cruzi

> **NIH NIH F32** · BROWN UNIVERSITY · 2024 · $74,284

## Abstract

PROJECT SUMMARY
Chagas' disease (CD) is a neglected tropical disease caused by the protozoan parasite Trypanosoma cruzi.
Over 70 million people in South and Central America, as well as immigrant populations, are at risk of CD. There
is currently no effective vaccine, and treatment options are limited. The parasite is transmitted by the feeding
activity of triatomine insects. Once in the blood stream of the mammalian host, the parasite invades nucleated
cells and proliferates until the cell ruptures, restarting the lytic infection cycle. T. cruzi has a unique morphology
with a single, anchored flagellum enclosed by the flagellar pocket (FP), and undergoes several morphological
changes during its life cycle that allow the parasite to adapt to the different environments within its mammalian
and insect hosts. The location of the FP and the flagellum along the cell body change during these transitions.
Of particular interest is the phase of the life cycle known as metacyclogenesis where insect forms of the parasite
(epimastigotes) differentiate into metacyclic trypomastigotes which are infectious to the mammalian host cell.
My goal is to identify novel molecular markers and describe the morphological changes that occur
during metacyclogenesis using transcriptomics and microscopy. In the first year of this fellowship, I will
focus on aim 1, where we will use single cell RNA sequencing to identify the gene cascades underlying
metacyclogenesis. From this, we will be able to identify molecular markers unique to either epimastigotes or
trypomastigotes, and be able to describe the gene regulatory cascades that underly both the epimastigote cell
cycle and metacyclogenesis. It is currently unknown if the transition from epimastigote to trypomastigote requires
a cell division, a question we will answer using live cell imaging. This will be facilitated by using fluorescently
tagged genes identified in the single cell transcriptomes to allow for real time imaging of transitioning parasites
to be validated by an expressed specific stage marker. At the end of the first year, and in the second and third
year, I will focus on aim 2 where we will map the localization of FP proteins during metacyclogenesis and
[validate their role in serum survival and host cell invasion efficiency]. In the former approach, we will use
CRISPR/Cas9 to endogenously tag several FP components and view transitioning cells using a combination of
conventional epifluorescence and ultrastructure expansion microscopy to generate high resolution structural
information. [Next, we will take FP mutants and expose them to complement active serum and measure lysis,
as well as allow them to invade host cell monolayers.] The findings of the proposed work will generate novel and
critical resources for the field of CD research, and reveal unique and essential biological pathways that may be
amenable [to transmission blocking] to combat CD. This work will be facilitated by the excellent research
environme...

## Key facts

- **NIH application ID:** 10998496
- **Project number:** 1F32AI181488-01A1
- **Recipient organization:** BROWN UNIVERSITY
- **Principal Investigator:** Caroline Dana Keroack
- **Activity code:** F32 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $74,284
- **Award type:** 1
- **Project period:** 2024-12-01 → 2027-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10998496

## Citation

> US National Institutes of Health, RePORTER application 10998496, Defining the molecular mediators of cell morphogenesis in Trypanosoma cruzi (1F32AI181488-01A1). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/10998496. Licensed CC0.

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