Murphy Lab Research
History and General Interests
Dr. Murphy began his research in the lab of Michael Sellix, Ph.D. as a laboratory technician following his B.S. in biomedical sciences from RIT. There he analyzed mammalian reproductive physiology in relation to the internal timing system in rodent models of hyperandrogenemia and targeted mammalian clock deletions in the ovary.
He later joined the lab of James Palis, M.D. at the University of Rochester as a Ph.D. candidate. There he investigated the cellular and molecular underpinnings of terminal erythroid cell maturation by determining the role of erythropoietin receptor in primitive erythroid cells (red blood cell precursors) and the associated downstream STAT3 and STAT5 signaling.
Dr. Murphy would then advance his studies as a postdoctoral associate in the lab of Laurie Steiner, M.D. where he investigated the epigenetic regulation of erythroid biology within the context of chromatin compaction and disease. Projects included the investigation of relevant chromatin regulators implicated in myelodysplastic syndrome, congenital dyserythropoietic anemia, and normal erythropoiesis.
Dr. Murphy is now continuing his work investigating the unique epigenetic environment that exists in erythrocyte (red blood cell) precursors. These cells have robust changes in cell size and chromatin organization, which when defective, lead to anemia. Anemia is any condition in which the body has a decreased ability to deliver oxygen. Anemia is the most prevalent condition worldwide affecting close to 4 billion people each year, with varying levels of severity.
Current projects are aimed at generating cell models of analysis of various anemias by using CRISPR-Cas9 in mammalian cell culture. Additionally, he is interested in comparing mechanisms of maturation between embryonic and adult hematopoiesis (blood cell formation).
Ongoing Projects
Generation of EPOR-independent HUDEP2 and KITCAT lines: The EPOR is a required receptor for erythroblast maturation. Using CRISPR Cas9 genome editing, the aim of this project is to create HUDEP2 and KITCAT cells lines that have mutations resulting in constitutively active EPOR. This will decrease cell culture costs but also serve as a model to study EPOR signaling in erythroblasts
Generation of a disease model for CDA-I: Congenital Dyserythropoietic Anemia Type I results from a mutation in CDAN1 which results in anemia. There are several mutations found in patients. The aim of this project is to create cell lines with patient mutations to mimic human disease. This project will lead into follow up analysis of mutant cell lines.
Determining if radioprotective mechanisms in murine erythroblasts are conserved in human cells: Congenital Dyserythropoietic Anemia Type I results from a mutation in CDAN1 which results in anemia. There are several mutations found in patients. The aim of this project is to create cell lines with patient mutations to mimic human disease. This project will lead into follow up analysis of mutant cell lines.
Investigation the role of TET2 in erythropoiesis: TET2 is a gene that is suggested to be related to MDS, and it may play a role in the proper functioning of erythropoiesis. Studies have shown the possibility of a lack of TET2 playing a role in the progression of MDS in patients. TET2’s function is currently unknown, but previous publications suggest that its function may be related to erythropoiesis.
Generating an In-Vitro Model of MPS IIIA “Sanfilippo Syndrome” Through Gene Editing: Autosomal recessive inheritance of a defective SGSH gene coding for the enzyme sulfamidase is responsible for mucopolysaccharidosis type IIIA, characterized by the effects of heparan sulfate accumulation in lysosomes. The syndrome presents with coarse facial features, central nervous system dysfunction, behavioral difficulties, intellectual disability, sleep disturbance, hyperactivity, loss of communication skills, and lack of mobility. Failure to develop cost effective research methods and long term treatment options leaves the average expectancy at just 15 years.
In vitro corneal wound healing model using HT-1080 fibrosarcoma cell lines: Our principal aim is to elucidate the effects of various ophthalmic solutions to test in vitro cellular proliferation, protein expression, and migration ability within a wound-healing model. This aim focuses on examining the effects different ophthalmic solutions have on epithelial cell lines. Epithelial cell lines such as A549s, ocular cell lines like human corneal epithelial cells (HCEC), or fibroblast-like cells will be used in this study. Our goal is to measure and determine the greatest growth rate and migratory ability by developing growth curves before and after treatments with various ophthalmic solutions. We will observe protein expression by Western blotting as a form of biochemical analysis. Moreover, to test cell migration ability, traditional cell scraping protocols will be followed. These tests will enable us to determine which ophthalmic solution provides the best wound-healing results.
Effects of glyphosate on human health: More research needs to be conducted to make direct conclusions regarding the correlation between cancerous outcomes and produce consumption containing glyphosate. A ‘safe’ limit for glyphosate on food consumed by humans considered by the EPA is “0.01 to 400 parts per million (ppm) and The Environmental Working Group (EWG) considers “160 parts per million the safe limit for human consumption” (alergyamulet.com). This is a lower range in comparison to the EPA standards. Glyphosate has been found in various crops including corn, soybean, oil seeds, grains and some fruits (fda.gov).
Potential Future Projects
Reverse Genetics; Determine phenotype of gene disruption in red blood cell precursors: Determine a gene that may be important for function in red blood cell precursors. Experiments would then disrupt this gene by CRISPR/Cas9 genome editing and analyze the phenotypic consequences in cells.
Optimization of homology directed repair following CRISPR/Cas9 editing: Use inhibitors of non-homologous end joining DNA repair in order increase efficiency of directed repairs in cells. Part of this project would also explore the potential of base change CRISPR/Cas9 in human cell culture.
Understanding the role of primary scientific literature on course performance: The use of primary scientific literature is more and more common in the classroom. However, these uses are often for specific content applications. The aim of this project is to determine if the practice of critical evaluation of primary literature can be used to enhance students' critical thinking and studying skills. This project would evaluate how students' perceptions on course content and the ability to adapt to that content are influenced by regular evaluation of literature. The hypothesis is that the critical thinking required for critical evaluation of primary literature benefits students’ ability to master content in courses in which there is no direct connection to the literature studied.
Determining if integration of formal scientific writing skills into a course benefits student outcomes: Many courses, particularly at the graduate level, require substantial writing. Often, student backgrounds are diverse and writing performance varies. The aim of this project is to determine if writing early intervention through direct course instruction increases student writing performance. Outcomes from this course will have a direct impact on the delivery of courses such as GPBH511.