Taufeeque Ali

Summary of Milwaukee Institute for Drug Discovery (MIDD) research assistantship

Goals of the research for Spring 2023:

1. Measure changes in tumor suppressor gene p53 mRNA levels and protein levels via RT-qPCR and Western blot.
2. Measure DNA damaging effect of prodrug, vitamin C and combinations in cells through Fluorescence Microscopy.
3. Determine microsomal stability and metabolites.
4. Determine the correlation between H₂O₂ level generated by vitamin C and the synergistic anticancer effect of the combination strategies.
5. Measure the Reactive Oxygen species (ROS) levels induced by vitamin C in vivo.
6. Determine the in vivo efficacy and selectivity of the combination strategies using xenograft models.
7. Determine Plasma pharmacokinetics and LC-MS/MS analysis.
8. Test the combination strategies in other cancer cell lines such as Glioblastoma brain tumor (Promising preliminary data).

I express gratitude to MIDD for granting this research assistantship, which has been instrumental in accomplishing diverse research objectives in Spring 2023. These goals encompass testing novel combination strategies in various cancer cell lines, establishing correlations between H₂O₂ levels and the anticancer activity of prodrugs, and evaluating associated DNA damage. The team, consisting of three undergraduate students with diverse academic backgrounds, actively participated in projects centered around ROS-generating agents, dose optimization, and unraveling the mechanisms of prodrug activation. Their collaborative endeavors have significantly contributed to this research project, leading to a patent application, securing a National Institutes of Health (NIH) grant and a catalyst grant, two publications and a review article under preparation. The research conducted in our lab aligns closely with their academic and professional aspirations.

Undergraduate student: Dhivyashree Senthil Murugan

I am Dhivyashree Senthil Murugan, a double major in Biological Sciences with a focus on neuroscience and Cell & Molecular Biology, and a minor in Chemistry. My passion for research led to the honor of receiving the SURF (Summer Undergraduate Research Fellowship) for both spring and summer 2022, along with the Senior Excellence in Research (SERA) award for the academic year 2022-23. Engaging in the L&S SS291 UROP Seminar and CHEM691 (Senior Research) over several semesters, I worked with graduate student Taufeeque Ali in Dr. Peng’s lab on the innovative project "Novel Combination Strategies for Tumor-Selective Activation of ROS-Activated DNA Cross-Linking Agents." This research broadened my understanding of natural ROS-generating agents and cancer types. Under Taufeeque's guidance, I focused on Indomethacin, a well-known ROS generator, exploring its cytotoxicity in various cell lines such as MDA-MB-468, MCF7, and U87. I learned techniques to investigate its effects on mRNA and protein levels through RT-qPCR and western blots. Our goal was to develop a novel combination strategy with a tumor-selective ROS-generating agent, Indomethacin and a ROS-activated prodrug developed in our lab. This experience not only enhanced my experimental skills but also provided opportunities to present our findings at research symposiums. My enthusiasm for developing targeted anticancer drugs aligns with my aspiration to attend medical school for an MD. I look forward to contributing to the field in the journey ahead.

Undergraduate student: Alexis Kimberly Peterson

As an undergraduate student majoring in Biomedical Sciences with a focus on medical lab work, I was honored to be selected as a recipient of the SURF (Summer Undergraduate Research Fellowship) in the fall of 2022 and the Senior Excellence in Research (SERA) award for the academic year 2023-24. I am currently immersed in CHEM691 (Senior Research). In the research project "Novel Combination Strategies for Tumor-Selective Activation of ROS-Activated DNA Cross-Linking Agents," my goals for Spring 2023 were focused on investigating the mechanism of H₂O₂ generation by prooxidants in both cancer cells and normal cells. Under the mentorship of Taufeeque in Dr. Peng's lab, I initiated my work with Vitamin C, delving into its effects on cancerous and normal cells. My findings played a crucial role in optimizing the prodrug and Vitamin C dose to achieve a synergistic effect that selectively eliminates cancer cells while sparing normal cells. This led to the quantification of H₂O₂ levels both extracellularly and intracellularly. Subsequently, I established the role of the H₂O₂-generator Vitamin C in prodrug activation, resulting in a 6-fold dose reduction compared to the prodrug alone. Encouraged by this evidence, I proceeded to conduct comet assays to further confirm the synergy between the two compounds. I have presented my work throughout UW campuses, and my work is ongoing. I continue to explore and work with more prooxidants. My goal is to carry forward this research as a master's student in Dr. Peng's lab. I am dedicated to the research process and anticipate that my contributions will greatly contribute to the project's success.

Undergraduate student: Jatin Pandey

My name is Jatin Pandey, and I am currently an undergraduate student pursuing a major in Microbiology with a minor in Chemistry. I had the honor of being selected as a recipient of the SURF (Summer Undergraduate Research Fellowship) in the fall of 2022. I am actively engaged in CHEM691 (Senior Research). My initial interest revolved around being involved in all aspects of Taufeeque's research in Dr. Peng’s lab. I was particularly drawn to Glioblastoma cancer due to its poor prognosis and urgent need for innovative therapies. However, my focus later shifted towards ROS quenchers as part of the biological investigation where our understanding of the prodrug's mechanism was limited. Given that the prodrugs are designed to activate in the presence of H₂O₂, my work centered on quenching this H₂O₂ and establishing its role in the drugs' selectivity towards cancer cells. I conducted various experiments to understand the endogenous antioxidative systems of both cancer and normal cells. With Taufeeque's guidance, I measured catalase activity in different cell lines, including MCF10A, MDA-MB-468, MCF7, and U87, along with their endogenous H₂O₂ levels. My overarching goal was to establish a correlation between H₂O₂ levels present in cells and enhanced prodrug activation in cancer cells compared to normal cells. Alongside my colleagues, I have contributed to obtaining scientific data for a patent application and existing NIH grant. My experiences and contributions in the laboratory align seamlessly with my aspirations to pursue a career in medicine.