2021 |
Parast, Mana M |
U54Activity Code Description: To support any part of the full range of research and development from very basic to clinical; may involve ancillary supportive activities such as protracted patient care necessary to the primary research or R&D effort. The spectrum of activities comprises a multidisciplinary attack on a specific disease entity or biomedical problem area. These differ from program project in that they are usually developed in response to an announcement of the programmatic needs of an Institute or Division and subsequently receive continuous attention from its staff. Centers may also serve as regional or national resources for special research purposes, with funding component staff helping to identify appropriate priority needs. |
3d Multiscale Spatial Mapping of the Human Placenta @ University of California, San Diego
SUMMARY The goal of OSP1 is to generate three-dimensional multiscale maps of the human placenta from healthy uncomplicated pregnancies. OSP1 will interact with the other FR TMC Cores/Projects and the other HuBMAP Centers to facilitate data and resource sharing across the Consortium and with the broader scientific community. The placenta is the interface between mother and fetus, mediating exchange of nutrients and metabolic wastes and producing endocrine signals that promote maintenance of the pregnancy and proper fetal growth. The placenta is comprised largely of cells of fetal origin, including stromal cells, capillary endothelial cells, and three types of trophoblast: proliferative cytotrophoblast; hormone-producing and transport-mediating syncytiotrophoblast; and invasive extravillous trophoblast. The placenta also contains fetal and maternal immune cells, which mediate immunologic responses to infection and may play roles in placental development. Abnormalities in placental development and function have been linked to the most common and serious complications of pregnancy, but details of the mechanisms leading to adverse pregnancy outcomes remain to be elucidated. To enable future studies aimed at identifying the structural and functional perturbations that underlie placental dysfunction-mediated pregnancy complications, we propose to generate a reference dataset from normal term placentas. Importantly, the complementary strengths of our investigative team enable us to obtain longitudinal prenatal in vivo MRI and ultrasound imaging data and post-delivery biomechanical and molecular profiling data from the same organs. Rigorous pre-analytical and characterization pipelines will ensure collection of high-quality biospecimens and generation of reproducible data. A range of advanced molecular profiling techniques will be used, including initial bulk and dissociated single-cell transcriptomic, chromatin accessibility, and extracellular matrix proteomic profiling to identify component cell types and prioritize targets. These targets will then be interrogated using high-resolution multiplexed spatial transcriptomic and imaging mass cytometry technologies. The resulting data, linked to comprehensive metadata, will be transferred on an ongoing basis to the DAC, and analyzed collaboratively with the DAC, and investigators at the HIVE. Finally, we will to generate 3D multiscale maps of the placenta that can be explored to gain novel insights into the physical and regulatory relationships among different cell types, between cells and their environment, and between tissue structure on the microscopic level and whole-organ function.
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0.958 |
2021 |
Parast, Mana M |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Human Trophoblast Stem Cells: the in Vivo Niche and Relationship to Pluripotent Stem Cells @ University of California, San Diego
Project Summary/Abstract The human placenta plays a major role in maintaining the proper environment for fetal growth, but remains as the most poorly understood organ. This project aims to substantially improve our knowledge of this important human organ by studying its early development in a systematic, detailed manner, then to combine this knowledge with the latest technologies in regenerative medicine in order to develop in vitro models for the study of both normal and abnormal placental development. Specifically, we aim to understand the mechanisms underlying the establishment and maintenance of a multipotent human trophoblast stem (TS) cell, one which can give rise to all other subtypes of trophoblast, the epithelial cells of the placenta. Over the past few years, we have identified a key pathway, directed by the p53-related protein, p63, which is required for maintenance of undifferentiated cytotrophoblast (CTB) stem cells in the early placenta. More recently, we have noted that a subset of these CTB co-express CDX2, a transcription factor required for maintenance of TS cells in mice, and hypothesize that these CDX2+/p63+ CTB are multipotent human TS cells. We will characterize this subpopulation further, using a combination of FACS sorting, followed by differentiation assays and both bulk and single cell transcriptome profiling. In this discovery-based approach, we will focus our analysis on identification of transcription factors and cell surface markers, which characterize this cell population in the early human placenta. At the same time, we will take a more gene-focused approach, probing the specific role(s) of p63 and CDX2 in first trimester CTB proliferation and differentiation, including their downstream targets. Finally, we will apply this knowledge to human pluripotent stem cells (hPSCs)--both embryonic (hESCs) and induced pluripotent stem cells (hiPSCs)?in order to develop in vitro models for the study of human trophoblast lineage specification and differentiation. We have established a novel differentiation protocol for step-wise differentiation of hPSCs, first into CTB, and subsequently into hCG- secreting syncytiotrophoblast (STB) and HLA-G+ extravillous trophoblast (EVT). Using this protocol, we have found that Trisomy 21 iPSC spend a prolonged period in the CTB stem cell state, and show blunted differentiation into functional STB, identical to the phenotype of primary CTB isolated from placentas with Trisomy 21. These exciting preliminary data suggest that hPSCs may be useful for modeling trophoblast differentiation defects, which are the basis for placental dysfunction. We will compare hPSC-derived trophoblast to primary trophoblast from both pre- and post-implantation tissues in order to determine which they most resemble. The successful completion of this project has the potential to transform the field of human placental biology, by both identifying human TS cells within the placenta, and establishing hPSC-based models of placental disease, thereby constructing a firm foundation on which diagnostic marker discovery and therapeutic targeting of this important human organ would be possible.
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0.958 |