Research Projects

DNA damage is a physiological process during placental development but an increase in such damage triggered by genetic predispositions, stress or environmental toxins, can severely affect placental development leading to pregnancy complications such as preeclampsia, fetal growth restrictions (FGR) and preterm birth. Mechanisms that regulate DNA damage in placental cells are largely unknown and understanding this process is crucial for elucidating its impact on pregnancy outcomes. Our long-term goal is to elucidate role of genome maintenance in placental development and its implications for pregnancy complications. These discoveries will facilitate understanding foundational knowledge of placental biology and aid in the design of novel diagnostics to predict pregnancy complications in early stages. The objective of this grant is to characterize cell free DNA (cfDNA) in mother’s circulation which are generated by DNA damage in placental cells. The central hypothesis is that placental cells are polyploid and undergoes multiple rounds of replication, leading to the accumulation of DNA damage at specific loci (known as fragile sites). These loci, specifically repetitive sequences are more sensitive for DNA damage in various pregnancy complications and this damage is likely to result in the release of cfDNA into maternal circulation. The rationale for this study is that identification of fragile DNA damage site will help in developing tools to detect pregnancy complications in early stages. To test the hypothesis our specific aims are: (Aim1) Identification of DNA damage sites in human placental cells and detection of associated genomic regions (Fragile DNA) in the serum of pregnant women. (Aim2) Measurement of levels of fragile DNA in various pregnancy complications with focus on pre-eclampsia/preterm birth/FGR. (Aim3) Creation of diagnostic tools to detect DNA breaks in early pregnancy. For aim 1, a novel protocol for mapping of DNA breaks in placental cells has been developed in mouse model in my lab. This tool will be used in human placental cells to map fragile DNA damage sites using human trophoblast stem cells and genomics approaches. Aim 2 will apply molecular approaches such as qPCR and digital droplet PCR (ddPCR) to detect target cfDNA in pregnancy complications. For aim 3, we will develop diagnostic tools to detect target cfDNA in pregnancy using CRISPR and lateral flow assay. This contribution is significant since it will provide critical insights into the mechanisms of pregnancy complications, facilitating early detection strategies that could significantly improve maternal and fetal health outcomes. The proposed research is innovative because we are using cutting-edge techniques and novel method to identify fragile genomic sites in placental cells and its early detection in pregnancy complication. The proposed research has potential to make substantial contribution to understanding of cfDNA role in diagnosing pregnancy complications.