RaMP Project Descriptions

Zakia Abdelhamed
MD & PhD

Human Genetics

Our previous publication documented the existence of the primary non motile cilia in the fallopian tube secretory epithelial cells (FTSECs). These cells were long considered as nonciliated and their main function is to secrete fluids, containing nutrients and growth factors, into fallopian tube lumen. The fallopian tube fluid secretion is to maintain the strictly controlled environment needed for gamete transport, maturation, fertilization, and earliest stages of embryonic development occur. Nothing is known about the role of the FTSECs primary cilia and the aim of this project and the RaMAP student will be to investigate this hypothesis in a primary cilia focused lab, the Stottmann laboratory. Our previous paper and report were described in human tissue and our previous RaMP student had investigated this in mouse and found that like humans, mice FTSECs carry primary cilia. The research plan is to conditionally knockout essential cilia genes in these cells using a FTSECs specific cre- recombinase mouse line for example the Pax8-rTTA-TTO-cre. The latter is an inducible cre recombinase in a tetracycline dependent manner under control of Pax8 promotor. The student researcher will contribute to mouse genetics, animal husbandry, induction of cre-recombination with exogenous injection of tetracycline at specific time points, histology of the mouse fallopian tubes, and immunohistochemically characterization of cilia on the FTSECs using confocal and highresolution radial fluctuation imaging of cilia.

In-person only
Days per week: 2-3 days
10/30 5pm Breakout: unavailable; email zakia.abdelhamed@cchmc.org for questions

Raquel Arroyo
PhD Postdoctoral Fellow

Neonatology and Pulmonary Biology

The work in our lab is focused on Surfactant Protein D (SP-D), a collectin protein which is part of the lung surfactant system. SP-D participates in the innate immune defense of the lungs where modulates the inflammatory response and it binds to invading pathogens, such as bacteria and viruses, and promotes their clearance from the lungs by phagocytic cells. SP-D is found in many other organs in the human body as well as it has been detected serum showing variable levels during the curse of certain pathologies, such as sepsis. However, the origin, presence and functional role of SP-D in some of these other locations is not well understood yet, such as serum SP-D in sepsis. The goal of this project is to study the extra-pulmonary role of surfactant protein SP-D in a multi-microbial sepsis model in rodents that exhibit a high systemic inflammatory response. The student will learn about the respiratory system and immunology, specially focused on the innate immune response and surfactant protein D (SP-D). A small sub-project will be assigned to the student accordingly to the time that they will expend in the lab. Supervision and teaching of the necessary techniques will be provided during that time. The project involves working with mice models, processing of samples in the lab including: ELISAs, Western Blots, quantification of inflammatory cells and microscopy. Work with recombinant proteins will be also performed. Results will be discussed with the mentor, encouraging the student to bring up their own ideas for the design of experiments and solutions to the problems faced at the same time than helping them to address those.

In-person only
2 days per week
10/30 5pm Breakout:

Madeline Bonfield
PhD Candidate

Neonatology and Pulmonary Biology

Our lab is interested in understanding the development of the innate immune system in newborns and how that developing immune system functions to protect neonates from disease. My project focuses on a particular cell of the innate immune compartment, group 3 innate lymphoid cells (ILC3s) and how their differentiation and function is coordinated within the developing lung. I am particularly interested in the role that the transcription factor, PPARy, plays in this process. We are working on developing a number of in vitro methods to test the role of PPARy in these cells, which a student researcher could be involved with. These methods include: 1. Testing pharmaceutical agonists and antagonists of PPARy activity on cultures ILCs isolated from murine tissues; 2. isolating cells for culture by processing murine tissue and purifying the cell types of interest; and 3. Using ELISA assay to determine the effects of PPARy expression on ILC3 function by quantifying cytokine production.

In-person or virtual only (depends upon project choice)
Days per week: 2-3 days if in-person
10/30 5pm Breakout: unavailable

Jennifer Bourn

Cancer Biology

Despite significant advances in early detection and treatment strategies, breast cancer remains a leading cause of cancer-related deaths among women in the United States, with an estimated 42,000 deaths in 2020 alone. To overcome these sobering statistics, the is an unmet need to elucidate novel molecular targets and therapeutic strategies for patients with advanced breast cancer. RON, a member of the Met receptor tyrosine kinase family, is expressed in greater than 50% of human breast cancers. More importantly, RON expression is associated with advanced disease, metastasis, and early death in breast cancer patients. We have shown that RON is a key driver of breast tumorigenesis and progression by promoting tumor cell survival, but the molecular mechanisms of how RON promotes tumor progression and immune suppression remains unknown. We have uncovered a novel association between RON and interleukin-1 receptor associated kinase (IRAK4) which results in the decreased activation of IRAK4. IRAK4 is a crucial signaling molecule involved in signaling innate immune responses from Toll-like receptors and is necessary to trigger release of pro-inflammatory mediators and innate immune actions. We also show that RON-mediated down regulation of IRAK4 is associated with a reduction in IRF7 activation, a transcription factor that serves as a master regulator of Type I interferons (IFN) production. Type l IFNs are released by cells and can act in an autocrine fashion to promote cell death or in a paracrine fashion to heighten immune responses. Based on these preliminary data, we propose to test the hypothesis that RON expression in breast cancer cells suppresses Type 1 IFN production through IRAK4 inhibition to promote aggressive tumor cell growth and immune cell evasion. The overarching goal of this study is to interrogate the extrinsic and intrinsic effects of RON expressing breast tumor cells on tumor growth and progression via the suppression of innate immune signaling. These studies will provide the scientific underpinning in the development of novel drug targets and therapeutic strategies crucial for the treatment of advanced breast cancer.

The role of the student researcher will be to aid the mentor in conducting the experiments required for this study but also develop and perform their own experiments to fulfill the requirements of the RaMP program.

The student will be trained in how to make the necessary reagents for the lab, learn and perform molecular biology techniques including but not limited to, DNA and RNA isolation, PCR, cell and tissue culture, western blot analysis, immunohistochemistry, microscopy, and animal handling.

In-person only
Days per week: 2-3 days; back-to-back days and larger time blocks preferred
10/30 5pm Breakout: unavailable; email bournjr@ucmail.uc.edu for questions

Nicole Edwards
Postdoctoral Fellow

Developmental Biology

Trachea-esophageal birth defects can occur when there are disruptions in the proper development of the trachea and esophagus. We are interested in determining if mutations in certain genes cause these developmental defects. In the Zorn lab, we are using frogs (Xenopus) and CRISPR-Cas9 gene editing technologies to screen for new gene mutations that we have discovered in human patients, and to see if mutating these genes in frogs also cause similar defects during frog embryo development.

The student will participate in this genetic screen by learning how to design CRISPR-Cas9 targeting strategies (can be done virtually), learn genotyping by PCR and sequencing, analyzing sequencing results (can be done virtually), immunofluorescent staining methods, and get hands on experience working with frog embryos.

In-person only
2-3 days per week, ideally back to back; 3-4hrs per day
10/30 5pm Breakout:

Demetria Fischesser
PhD, Postdoctoral Fellow

Department of Internal Medicine, Cardiology Division

It is well known that myocardial infarctions (heart attacks) cause a substantial amount of cell death in the heart. However, upon removal of the infarction-inducing blockage, re-introduction of oxygen to the tissue causes a second wave of cell death. It has been hypothesized that tempering this oxygen reintroduction could have a positive therapeutic benefit for patients. In order to investigate this hypothesis, the Haworth lab is currently using ultrasound cavitation to disrupt perfused droplets containing an oxygen scavenger.

A major aspect of this project involves the generation of these droplets using microfluidics. This process allows us to synthesize droplets of various sizes (1um-12um) which contain an oxygenscavenging material, perfluoropentane.

However, we want to explore new possibilities for generation of these oxygen-scavenging droplets by altering their synthesis and makeup. The student will be primarily responsible for testing how different materials and synthesis methods affect the size and oxygen-scavenging abilities of these microfluidics droplets.

In-person only
3-4 days per week
10/30 5pm Breakout: unavailable; email michaedt@mail.uc.edu with any questions

Chenna Galiveti
PhD, Senior Postdoctoral Fellow

Department of Environmental & Public Health Sciences

Under the tutelage of Dr. Galiveti, the student will culture head and neck cancer cells and primary oral epithelial cells and harvest small extracellular vesicles ("exosomes") from conditioned culture media. Exosomes will be fixed, stained with fluorescent antibodies against candidate cytosolic membrane-associated proteins and imaged via super resolution microscopy (CCHMC Confocal Imaging Core).

The student will be responsible for performing cell culture, small EV isolation, nanoparticle tracking analysis, and fluorescent staining under the supervision and mentorship of Dr. Galiveti.

In-person primarily, with a few virtual opportunities
Days per week: 2-3 days per week; 10-15 hours but flexible in both
10/30 5pm Breakout:
https://ucincinnati.webex.com/ucincinnati/j.php?MTID=mcad8fd708320037ad012ca0396ca013 5
11/2 4pm Breakout offered as well:
https://ucincinnati.webex.com/ucincinnati/j.php?MTID=m2a83c6964200f6868d28379396f9c3a f

Tushar Ganjawala

Developmental Biology

Abstract: Wnt signaling plays a critical role in development and diverse cellular functions, and so as its mis-regulation has been found to be linked with several developmental diseases as well as cancers. Within a developing embryo, Wnt signal is known to cause distinctive effect in different cell types simultaneously, called context dependency, through co-regulation by differentially targeting diverse transcription factors. Our lab studies such transcriptional regulators, cis-regulatory elements, associated with Wnt signaling pathways using a simple animal model system, the nematode worm C. elegans. We use computational methods to predict new target genes/gene sequences and then apply molecular biology techniques and confocal microscopy to test them in vivo in C. elegans embryo. The invariant lineage of C. elegans enables a time-lapse imaging approach to trace expression of a gene of interest in wildtype or mutant embryos. Specifically, we propose to explore how nhr-67 enhancers are influenced by Wnt signaling in context dependent manner.

Mentor’s responsibility: As a mentor, I commit to explain basics of relevant research methods and goals and assist a student researcher performing them efficiently and confidently. Help student achieve his/her goal in time. Student’s responsibility: Student will learn the concept of relevant research methods and expected to do following with some assistance as and when required;

  1. Some literature review to get background knowledge 
  2. Use computational methods to find and propose TF sites 
  3. Use molecular biology techniques to clone and generate mutant and synthetic enhancers 
  4. Use microscopy to see and evaluate fluorescent reporters in C. elegans

Student may work virtually for some computational methods. Also, if time permits, student may learn evaluating time-lapse images (movie) using software. At the end of the project student will have data to present his/her work in form of a poster or a short talk.

In-person only
Days per week: 3-4 days; 12 hours per week
10/30 5pm Breakout: unavailable; email Tushar.Ganjawala@cchmc.org with questions

Marie-Eve Hoeppli


Children with Complex Regional Pain Syndrome CRPS) require outpatient and sometimes inpatient intensive treatment. The goal of this study is to define markers that predict CRPS recovery with outpatient treatment or with inpatient treatment. Definition of these predictors is essential to improve individualized treatment for these children. We are currently trying to standardize and improve the safety of the experimenter and participants during testing. The main role of the student working with us on this study will be to help with this standardization. In particular, this student will be responsible for the 3D design of support hardware for testing probes that will then be 3D printed in the lab.

Virtual, with a small number of in-person times necessary
Days per week: flexible; 12 hours per week
10/30 5pm Breakout:

Katherine Inskeep
PhD Student

Human Genetics/ Developmental Biology

I am studying a gene called Smpd4 which was found to cause microcephaly (small brain), epilepsy, and various other developmental brain effects in human patients. Part of my project is modeling this gene in mice. The student would assist with the Smpd4;Emx1-Cre mice, a line which has the Smpd4 gene deleted only in the brain. Their job would be to identify mutant mice via PCR/gel electrophoresis and then characterize their brains to look for abnormalities, using paraffin wax embedding, H&E staining, and brain measurements on a microscope. They may be involved in other projects as well, depending on the status of these mice.

In-person only Days per week: flexible (8-12 hours per week total) 10/30 5pm Breakout: https://us02web.zoom.us/j/87692406948?pwd=YWlTV3FwR2NFTXlOa2NvajRNSEFzUT09