Dr. Mei Lin Chan’s Osteoimmunology Lab (April 2017 – present):
Low Intensity Vibration May Accelerate CAR T Cell Biomanufacturing for Cancer Immunotherapy (January 2020 – October 2020)
Problem:
CAR T cell, or chimeric antigen receptor T cell, therapy is an autologous cell-based therapy at the cutting edge of immunotherapies against cancer. The current pipeline for CAR T cell manufacturing lasts 4 weeks from start to finish, with a substantial fraction spent on the ex vivo cell expansion phase (i.e., 2 weeks) alone, using IL-2, a potent T cell growth factor, and antigen presenting cell (APC) mimicking agents. To meet the time-sensitive demands of cancer treatment with a non-chemical approach, we propose a novel means to expedite the expansion phase for high-throughput CAR T cell manufacturing using mechanical stimulations. Preliminary studies have demonstrated the proliferative effects of mechanical signals delivered via low intensity vibration (LIV) in T cells. With a proprietary LIV protocol developed in our lab, the goal of this experiment is two-fold: to further validate preliminary results of rapid T cell expansion with the hypothesis that LIV-induced proliferation (i) will increase cell number and (ii) will not negatively alter cell biomarker expression and subset populations.
Role:
As a research assistant, I was responsible for conducting cell culture studies of CD3+ Pan T cells under BSL-2 cGLP laboratory conditions, planning experiments, and performing data analysis. Here I gained further experience with techniques such as flow cytometry, hemocytometry, and 2-photon microscopy. I took on a larger leadership role within the lab where I mentored several undergraduate students, which helped me to better understand and communicate my research.
Presentation(s):
Biomedical Engineering Society (BMES) Annual Meeting 2020 – Virtual Platform Presentation
URECA (Undergraduate Research & Creative Activities) Symposium Poster Presentation (May 2020)
Validation of Low Intensity Vibration-Induced T Cell Proliferation: A Story of Cell Number and Subset Population (October 2018 – October 2019)
Problem:
The current pipeline for CAR T cell manufacturing lasts 4 weeks from start to finish, with a substantial fraction spent on the cell expansion phase (i.e., 2 weeks). To meet the time-sensitive demands of cancer treatment, a new biotechnology solution is needed to expedite the expansion phase for high-throughput CAR T cell manufacturing. Refractory period (RP) is the resting period between low intensity vibration (LIV) bouts. With a patent-pending LIV protocol developed in our lab, the goal of this experiment is two-fold: (i) to validate our preliminary results of rapid T cell expansion for potential use in expedited CAR T cell manufacturing with the hypothesis that LIV-induced proliferation will increase rates of DNA synthesis with no compromise to viability and (ii) to investigate the validity of the current RP to maximize the proliferation response.
Role:
Here I was responsible for rapidly learning and applying various advanced cell culture techniques for CD3+ Pan T cells and CD4+ or CD8+ subtypes such as flow cytometry. I performed a multitude of experiments with LIV to determine the optimal parameters needed for increased cell proliferation. I used a novel cell proliferation/DNA synthesis assay to validate these results at the biochemical level.
Presentation(s):
Biomedical Engineering Society (BMES) Annual Meeting 2019 – Poster Presentation (October 2019)
URECA (Undergraduate Research & Creative Activities) Symposium Poster Presentation (May 2020)
Custom Image Analysis Algorithm for Analysis of Cytoskeleton and Morphology of Pre-adipocytes (August 2017 – December 2018)
Problem:
As the major cellular organelles of neutral lipids, lipid droplets (LD) serve an essential role in lipid metabolism and intracellular lipid storage regulation. Thus, they are important in the development of prevalent metabolic diseases such as obesity, diabetes and atherosclerosis. Our experiment aims to observe how adipocytes respond towards mechanical stimulation, especially how their number, size and distribution are regulated. This will improve our understanding of metabolic and potential molecular mechanism of adipocytes. The objective of our project is to develop, and validate an algorithm to analyze if the low intensity vibration (LIV) increases actin content, and changes morphology of preadipocytes. Specifically, to obtain image intensity, and morphological parameters (area within boundary, lengths of longest and shortest axes).
Role:
Here I applied my programming skills to generate a MATLAB based image analysis algorithm to identify morphological changes in adipocytes in response to mechanical stimulation. I was simultaneously responsible for learning preliminary cell culture techniques for adherent cell lines (3T3-L1 fibroblasts – murine adipocytes). I primarily learned various fluorescence imaging techniques such as 2-photon microscopy. We also performed various cytoskeletal knockout studies to determine the mechanism of morphological changes in response to LIV.
Presentation(s):
URECA (Undergraduate Research & Creative Activities) Symposium Poster Presentation (May 2019)
A Family-Based Fitness Wearable for Monitoring Childhood Obesity (April 2017 – August 2018)
Problem:
Childhood obesity is a growing issue in the United States. Obesity can be the cause of
many life-threatening diseases and can affect a person’s overall quality of life. While diet and exercise is the typical prescription for obesity, studies show that family involvement is often crucial in the treatment of childhood obesity. In fact, children with active parents are 5.8 times more likely to be active than children with inactive parents. Recent programs like Fit Kids for Life! encourage parents and children to exercise together and teach families how to incorporate exercise into their everyday lives. According to the Center for Disease Control, children should engage in 60 minutes of moderate or vigorous physical activity daily, with most of this activity being aerobic activities. The overall goals of the development are to effectively display and interpret the data obtained and safely secure the electronics in a durable yet comfortable housing for the user to wear. The accomplishment of these goals will result in a family-based fitness wearable for improving childhood obesity.
Role:
To quantify and improve family’s involvement, our project aims to develop a fitness wearable that encourages parents and children to exercise together through an interactive, competitive video game which uses heart rate monitoring technology to earn points and Bluetooth technology to connect the parent’s and child’s devices. In this project, the heart rate sensor and accelerometer are used to measure the user’s heart rate and number of steps as the key indicators of their level of physical activity. This device prototype is made wearable by the custom-designed housing and band made from Ninjaflex 3D printed filament.
Presentation(s):
URECA (Undergraduate Research & Creative Activities) Symposium Poster Presentation (May 2019)
Dr. Richard A Clark’s Dermatology/Tissue Engineering Lab (April 2017 – May 2020):
Preclinical cp12 Infusion Burn Studies (April 2017 – May 2020)
Problem:
Burn wounds of 2nd degree or greater can result in permanent scarring leading to a severe decrease in patient quality of life (diminished sweat production, decreased skin elasticity, location dependent inhibited mobility, etc.). This is because the first 24 hours are critical for burn injury progression, and thus an intervention within this time period to halt progression and promote healing would have great clinical relevance in improving burn patient outcomes.
Role:
As a research assistant, I was responsible for veterinary care of porcine subjects in the Division of Laboratory Animal Resources (DLAR), as well as performing whole body autopsies and histological tissue analysis. Under IRB and DOD approved protocols I helped to administer a novel fibronectin fragment based peptide drug, cp12, intravenously 2-6 hours post-burn injury for safety and efficacy preclinical studies in porcine subjects. Here I also developed a MATLAB image analysis algorithm to determine burn depth from non-invasive near infrared FLIR images of wounds within the critical first 2 weeks post-injury.
Preclinical cp12 Mesenchymal Stem Cell In Vitro Studies (May 2019 – May 2020)
Problem:
The exact pleiotropic mechanisms of action for cp12-induced accelerated burn wound healing are not well understood. One hypothesis is the influence on the shifting of the production of the cytokine panel from a pro-inflammatory state to a pro-wound-healing state.
Role:
As a research assistant, I was responsible for maintaining MSC fibroblast cells under BSL-2 conditions for various experiments including in vitro assays for cytokine production. I was instrumental in performing ELISAs for Type 1 collagen assays to reveal cp12’s induction of the formation of granulation tissue of wound contraction and the induction of epithelialization. I learned many new cell culture and data analysis techniques from this project.
Senior Design – Dr. Daniel Birk & Dr. Wei Rubenstein
Apto: Adaptors for Surgical Retractors
Problem:
Self-retaining surgical retractors are surgical instruments used to separate the edges of an
incision or hold back underlying tissues to enhance visibility and access for the surgeon. While these retractors grant surgeons the exposure needed, the current model exerts significant non-uniform force on the tissues they retract and requires surgeons to loosen retraction throughout the surgery, minimizing surgical exposure. The minimization of surgical exposure causes low visibility for the surgeon and limits the surgeon’s ability to properly perform the surgery. If the retractors are not loosened, however, this can cause irreversible damage to the patient’s tissue and result in unintentional ischemia, bleeding, necrosis, and chronic pain, resulting in extreme health and financial consequences for the patient.
To solve this problem, we propose Apto: slide-on, stationary retractor adaptors that
create a uniform force distribution on retracted tissues. These adaptors directly attach to the retractor, replacing the damaging sharp prongs with softer grips to grip the tissue. Apto retractor adaptors provide an effective solution for minimizing tissue damage due to retraction while maintaining the ease of use of the retractor. Our main objectives are to test the effectiveness of Apto with theoretical modeling and qualitative analysis, and to optimize Apto’s design. We seek to label one of the predetermined Apto designs as most effective and further optimize this design.
Role:
Here my role was the experimentalist where my responsibilities included planning ex vivo animal experiments, biomechanical tests, theoretical analyses for eventual translation into use for human tissue retraction. I helped to organize and order materials, plan experiments, collect data, provide reports and create figures from our various analytical techniques. I helped facilitate the collaboration between the Stony Brook Hospital, the Department of Biomedical Engineering, and the Department of Physics for the execution of these pre-clinical biomechanical studies.
Presentation(s):
URECA (Undergraduate Research & Creative Activities) Symposium Poster Presentation (May 2020)
Honors College Senior Symposium 2020 (May 2020)
Stony Brook University iCREATE WolfieTank 2019 (November 2019)
