2024 Project Descriptions
Project One
Elizabeth Wellberg, Ph.D.
Assistant Professor
elizabeth-wellberg@ouhsc.edu
Description:
Dr. Wellberg previously did a study that looked at high-fat diet (HFD) in a transgenic mouse model of HER2+ breast cancer. The HFD accelerated tumor formation compared to low-fat diet-all mice are done and Kaplan Meier data are significantly different.
HFD mice had more body fat, more insulin, more glucose, and probably other systemic issues. We banked a bunch of tissues including tumors and "normal" mammary glands, which have a mixture of normal cells and very small early DCIS-looking lesions. We've stained all of those tissues with a panel of ~8 antibodies to screen the infiltrating immune cells-data analysis is in progress. We have also already done Ki67 on the tissues and have images scanned into Aperio for analysis.
Things that need to be done as part of this project:
- p63/SMA on all tissues and lesions classified as in situ or invasive-many are likely to be invasive-we have the antibodes and SMA is optimized, p63 needs another round of optimization but we're close.
- region selection and algorithms run on each Ki67 stained lesion in Aperio and then data need to be looked at different ways to see if the proliferation correlates with any parameter such as tumor size, immune cell infiltration, and if that's related to diet, insulin, body fat % etc. (This would be very computer-centric type of work)-could be combined with TUNEL staining and analysis if there is time/interest.
- Any follow-up IHC on the tissues for new markers, hypotheses, etc-for example we have a lot going on with YAP and ETV4 transcription factors, tumor metabolic changes, ECM deposition with methods that are already optimized or close to it. Any of these could be interrogated in this sample set.
Project Two
Title: Cystathionine beta-synthase: Driver of multicellular crosstalk in the ovarian tumor microenvironment
Geeta Rao (PhD)
Assistant Professor,
Department of Pathology
P: 405-271- 8001 Ext. 58012
Email: Geeta-rao@ouhsc.edu
Abstract: Ovarian cancer (OvCa) is one of the deadliest gynecological malignancies. Emerging evidence suggests that the tumor microenvironment (TME) plays a critical role in the poor prognosis of OvCa, including high-grade serous OvCa (HGSOC), the gynecologic malignancy with the lowest survival rates. Cancer-associated fibroblasts (CAFs), cancer cells (CCs), and endothelial cells (ECs) are the key components of the TME, and crosstalk between them plays a central role in all stages of OvCa development and progression. However, we lack fundamental information about the regulator(s) of multicellular crosstalk within the TME which leads to poor prognosis and drug resistance. Recently, our group demonstrated the role of the metabolic enzyme cystathionine beta-synthase (CBS) in OvCa progression, therapy resistance, enhanced lipogenesis, and mitochondrial dynamics as well as EC bioenergetics and mitochondrial dynamics. Notably, previous reports suggest that CBS is secreted by microvascular ECs.Given the role of CBS in OvCa progression, and EC function, and since CBS is secreted by microvascular ECs, we hypothesize that CBS is a key enzyme regulator of multicellular crosstalk in the OvCa TME.
In this summer research project, we will test our hypothesis by disrupting multicellular crosstalk in the OvCa TME via CBS silencing. To unravel the specific contributions of CBS in distinct cell types (CC, CAF, or EC), we will use siRNA-mediated CBS silencing in each cell type. In this approach, cells will be plated in a ratio of 1:1:1, and we will evaluate the cellular phenotypes associated with each cell type. For ECs, we will assess tube formation and proliferation as key indicators. In the case of CAF, our analysis will encompass proliferation assays and the quantification of α-SMA and FAP expression levels. For CC, proliferation and AlexaFluor 568 labeled gelatin-invasion assays will be performed. Additionally, we will also measure CBS levels using immunoblotting and immunocytochemistry. We expect that these measurements will correlate with alterations in cellular phenotypes resulting from CBS inhibition and enable us to dissect the intricate interplay between CBS and multicellular crosstalk in the OvCa TME.
Project Three