||Women usually live longer than men and this longevity in women is likely due to the ability of women to conceive child. Pregnancy is not merely a childbearing experience, as it promotes a wonderful physiological alteration in the pregnant mother. In this regard, during pregnancy the fetal cells (FCs) have been shown to migrate from developing fetus, into the maternal circulation, and these FCs are known to remain for decades in maternal circulation. However, the exact role of these FCs are not completely understood. Therefore, my research project will be to characterize the phenotype of these FCs at the cellular and molecular level using lineage tracing genetic technology in the mice. Thereafter, I will address the ability of these FCs to repair tissue damage and restore organ function in the aftermath of experimental tissue injury such as ARDS, acute myocardial infarction (AMI), and peripheral arterial disease (PAD).
||Kishore Wary, PhD and Richard Minshall, PhD
||Butler, Mitchel "Mitch"
||Subarachnoid hemorrhage (SAH) is a debilitating injury most commonly caused by ruptured cerebral aneurysm; the disease leads to significant functional neurologic disabilities, including epilepsy. Prior preclinical and clinical studies have revealed associations between SAH-induced vascular damage and altered inflammatory as well as electrical signaling mechanisms in the brain. Our lab combines the endovascular perforation rat model of SAH with long-term video-EEG monitoring together with downstream measurements of vascular changes and neuroinflammation, specifically increases in microglia. In parallel, we leverage longitudinal clinical, EEG, and computed tomography (CT) brain imaging data to investigate how SAH injury impacts electrical activity and epileptic outcomes in the human. In this combined preclinical and clinical project, I am developing imaging-based methods to (1) quantify changes in the extent and location of bleeding in the brain and (2) relate them to metrics of electrical and neuroinflammatory activity.
||Jeffrey Loeb, MD, PhD and Jan Kitajewski, PhD
||Ewenighi-Amankwah, Chinwe, PhD
||Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer characterized by lack of expression of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 (HER2) overexpression. TNBC is known for its aggressiveness, organ metastases, and poor prognosis than other types of breast cancer. TNBCs are more common among African-ancestry populations. TNBC accounts for 39% of breast cancers in African American women under the age of 50, but only 16% in Caucasian women of the same age group. Because TNBC lacks receptors for estrogen, progesterone, and HER2 overexpression, there are no targeted therapies, and affected patients rely only on chemotherapy. Notch4 from a family of the Notch signaling pathway plays crucial roles in cellular developmental pathways, including proliferation, differentiation, and apoptosis. Of all four Notch receptors, Notch4 is more attractive in TNBC. A positive correlation exists between TNBCs and high expression of Notch4. There is a need for targeted therapy for TNBC, and notch4 seems promising. My study investigates the role of Notch4 in tumor endothelium, growth, vascularization, metastasis, and therapeutic targeting.
||Jan Kitajewski, PhD and Kishore Wary, PhD
||Triple negative breast cancer (TNBC) makes up to 20% of breast cancer diagnoses, with a 5-year survival as low as 11% for late stage metastatic disease. Unfortunately, while surgery and adjuvant chemotherapy are effective treatment option for early stage disease, there are very few therapeutic options for advanced metastatic TNBC spread. Importantly, TNBC is unresponsive to hormonal and targeted therapies of other types of breast cancer. In the complex metastatic cascade, Circulating Tumor Cells (CTCs) cross the endothelial barrier twice: first to enter systemic circulation (intravasation) and then to exit circulation (extravasation). My proposal aims to study Jag1-mediated Notch signaling as a potential target to limit TNBC metastatic spread by focusing on tumor-endothelial interactions during tumor cell extravasation from circulation into secondary organs. Our previous work targeting Jag1- mediated Notch signaling with Notch1 decoys attenuated TNBC endothelial binding and transendothelial migration (TEM). We therefore hypothesize that Jag1 is a novel mediator of TNBC extravasation. Theoretically, Jag1 could activate endothelial or tumor Notch, as the receptor is expressed on both cell types. To address both of these possibilities, I have generated CRISPR/Cas9 treated TNBC clonal cell lines targeting Jag1. To address intrinsic tumor Notch signaling, I will engineer TNBC cell lines with a dominant-negative inhibitor of intrinsic Notch signaling. Using these tools, extravasation will be modelled in vitro, in a cutting-edge microfluidics system that recapitulates the capillary microenvironment, and in vivo via intravenous injection of TNBC cells followed by tracking of lung capillary extravasation. The goal of my research proposal is to interrogate a Notch-based mechanism of TNBC extravasation during metastasis.
||Jan Kitajewski, PhD and Jalees Rehman, PhD
||McCann, Maximilian A, PhD
||The integrity of the endothelial barrier in the retina is critical, as disruption of this barrier leads to vessel leakage, macular swelling, and visual impairments in patients with diabetic retinopathy. Vascular endothelial growth factor (VEGF) is a key inducer of endothelial barrier permeability and is chronically elevated in many patients with diabetes. It acts in two phases. In the initial phase, VEGF induces signaling events that physically open the barrier, allowing vessel leakage. The second phase is less defined and brings about a prolonged effect that has a slower onset, which suggests transcriptional changes are at play. Neutralization of VEGF with anti-VEGF therapies has proven to be a vital treatment that lowers VEGF levels, and leads to reclosure of breached endothelial barriers, resolves macular edema, and restores vision. However, not all patients benefit from anti-VEGF, and even those who initially respond well can become unresponsive. Therefore, alternative therapies to anti-VEGF are needed. My research focuses on laying the groundwork for these therapies by developing a better understanding of the processes that govern pathological vessel opening and reclosure.
||Jan Kitajewski, PhD