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Cellular and Molecular Bioengineering

Cellular and Molecular Bioengineering - Poster Session E

Poster I1 - Synthetic Biology Applications in Diagnosing and Treating Preeclampsia

Saturday, October 26, 2024

10:00 AM – 11:00 AM EST

Location: Exhibit Hall E, F & G

Presenting Author(s)

MD

Madeline Dunsmore

Student
Rowan University
Sewell, New Jersey, United States

Co-Author(s)

LD

Leah Davis

Ph D
Rowan University, United States

Primary Investigator(s)

Mary Staehle, PhD (she/her/hers)

Department Head and Professor
Rowan University, United States

Introduction: Preeclampsia ranks among the top three causes of maternal morbidity and mortality. Recent research indicates that placental hypoxia plays a crucial role in triggering the inflammatory response responsible for this life-threatening complication in both the mother and fetus. Current diagnostic methods for preeclampsia rely on detecting late-stage symptoms such as high blood pressure and proteinuria. Therefore, developing tools for real-time monitoring of placental oxygenation could significantly enhance early prediction and prevention of preeclampsia. This study focuses on creating a synthetic phosphorylation network that activates in low-oxygen environments, aiming to improve the diagnosis and treatment of preeclampsia. Synthetic transmembrane receptors have demonstrated their potential in regulating cellular responses under specific conditions, and thus for the basis for our synthetic phosphorylation network.

Materials and

Methods: To develop the biosensor, we constructed a post-translational circuit (PTC) that includes an extracellular binding domain, a transmembrane domain, and an intracellular signaling domain. When the extracellular portion dimerizes, it triggers an intracellular phosphorylation cascade that can lead to various cellular responses. We integrated oxygen-dependent degradation domains of HIF-1α and HIF-2α into the PTC. Activation of the biosensor initiated a unique phosphorylation cascade that resulted in reconstitution of a split fluorescent protein. HEK-293T cells were transfected using chemical transient transfection and then exposed to both normoxic (21%) and hypoxic (1%) conditions. Flow cytometry was employed to measure the Mean Fluorescent Intensity (MFI) of mNeonGreen, serving as an indicator of expression

Results, Conclusions, and Discussions: The engineered phosphorylation network, incorporating HIF-1α and HIF-2α, selectively activated under hypoxic conditions. Both HIF-1α and HIF-2α showed higher activation at 1% oxygen compared to 21% oxygen. Notably, HIF-2α exhibited a greater increase in activation under hypoxic conditions than HIF-1α, highlighting their distinct roles in the hypoxic response. The differential activation of HIF-1α and HIF-2α in chronic hypoxic conditions underscores their unique roles in regulating the hypoxic response. These findings suggest that different hypoxia-inducible elements may be involved in various stages of preeclampsia development through placental hypoxia. Taken together, this study provides insights into how specific hypoxia-inducible elements could correlate with the progression of preeclampsia. Developing methods to deploy this engineered phosphorylation network could significantly enhance real-time measurement of cellular hypoxic responses. Early detection of hypoxic conditions could prevent birth defects arising from placental hypoxia. Additionally, this technology could lead to targeted therapies addressing improper spiral artery remodeling, a primary cause of placental hypoxia and preeclampsia, thereby improving maternal and fetal health outcomes.

Acknowledgements (Optional):