Investigate the effects of rare damaging mutations in TGFB latent complex proteins found in preeclamptic placentas

Project Summary/Abstract
Preeclampsia (PE) is a severe pregnancy complication that affects 8% of pregnancies globally,
contributing to over 50,000 maternal and 500,000 fetal deaths annually1. Characterized by hypertension, PE
arises from poor placental development, partially due to impaired extravillous trophoblast (EVT) differentiation
and invasion. Transforming growth factor-beta (TGFβ) signaling is essential for trophoblast differentiation and
function, regulating cell migration and extracellular matrix (ECM) remodeling4. Dysregulated TGFβ signaling,
including elevated TGFβ levels in preeclamptic placentas, disrupts these processes, leading to poor placental
function and adverse outcomes1.
Our research focuses on rare damaging mutations in key TGFβ latent complex proteins, including
Fibronectin 1 (FN1) and Latent Transforming Growth Factor Beta Binding Protein 1 (LTBP1), that we identified
in placentas and umbilical cord blood mesenchymal stem cells (MSCs) from PE-affected pregnancies. We
hypothesize that these mutations impair TGFβ latency, enhancing active TGFβ signaling, and disrupting ECM
integrity which could ultimately affect EVT migration, invasion, and differentiation. Using patient-derived cells
and CRISPR-engineered models, we aim to investigate the functional consequences of these mutations on
TGFβ production, signaling, and downstream molecular pathways.
In Aim 1, we will study the effects of TGFβ activation and signaling in UC-MSC and iPSC-derived
trophoblast stem cells (TSCs) from PE-affected and healthy placentas. We will analyze TGFβ levels, TGFβ down
stream signaling, EVT differentiation, and perform functional assays to measure migration and invasion. Bulk
RNA sequencing will uncover dysregulated pathways contributing to PE. In Aim 2, we will use CRISPR/Cas9
technology to introduce specific mutations into iPSCs, differentiate them into TSCs and EVTs, and test their
effects on ECM integrity, TGFβ latency, and EVT function. RNA sequencing will further identify mutation-specific
molecular disruptions.
The results have the potential to inform new diagnostic tools, therapeutic targets, and preventive
strategies for PE, ultimately improving maternal and fetal health outcomes. Additionally, this research may
extend to other placental disorders involving TGFβ signaling and ECM dysfunction, contributing to a broader
understanding of placental development and disease.

Grant Number: 1F32HD120014-01
NIH Institute/Center: NIH
Principal Investigator: Cindy Barba