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

The National Institute of Child Health and Human Development awarded the University of California, San Diego $230,388 on January 25, 2026, under the Child Health and Human Development Extramural Research program (CFDA 93.865) to investigate the effects of rare damaging mutations in transforming growth factor-beta (TGFB) latent complex proteins found in preeclamptic placentas.

The research examines how mutations in fibronectin 1 (FN1) and latent transforming growth factor beta binding protein 1 (LTBP1)—identified in placentas and umbilical cord blood mesenchymal stem cells from preeclampsia-affected pregnancies—impair TGFB latency and disrupt extracellular matrix integrity. The investigator hypothesizes these mutations enhance active TGFB signaling and ultimately affect extravillous trophoblast migration, invasion, and differentiation, contributing to poor placental development.

The project uses patient-derived cells and CRISPR-engineered models to investigate functional consequences of the mutations on TGFB production, signaling, and downstream molecular pathways. Aim 1 analyzes TGFB activation and signaling in umbilical cord mesenchymal stem cells and induced pluripotent stem cell-derived trophoblast stem cells from affected and healthy placentas, performing functional assays to measure migration and invasion and bulk RNA sequencing to identify dysregulated pathways. Aim 2 employs CRISPR/Cas9 technology to introduce specific mutations into induced pluripotent stem cells and differentiate them into trophoblast models.

Work is performed in La Jolla, California with a period of performance through July 31, 2027.

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.