Topic
Human Rights and Political Determinants of Health

URCF is pleased to announce our Spring 2024 workshop schedules covering topics on Linux, Research Computing, and Introduction to Programming in Python and R. The workshops are disseminated live via Zoom.

Our training schedule and registration link for the Spring 2024 quarter is as follows:

Drexel Orthopaedic Research Day is open to all students interested in learning more about orthopaedic research or who have/are participating in current orthopeadic research! This half-day event will encompass multiple DUCOM alumni guest speakers and opportunities for students to share their research experiences. We invite students to submit abstracts prior to the event to be selected for either a podium or E-poster presentation to share their research. Following orthopaedic specialty lectures in the morning, we will host two E-poster sessions in the 4th floor labs and selected podium presentations of students' work. Both the poster sessions and podium presentations will be judged by DUCOM alumni in the field with all students that present being eligible for award recognition.

BIOMED PhD Thesis Defense

Title:
Type V Collagen Orchestrates the Initial Matrix Templating in Developing Articular Cartilage and Meniscus

Speaker:
Bryan Kwok, PhD Candidate
School of Biomedical Engineering, Science and Health Systems
Drexel University
 
Advisor:
Lin Han, PhD
Professor
School of Biomedical Engineering, Science and Health Systems
Drexel University

Details:
The articular cartilage and meniscus in the knee joint are critical for painless movement, but these tissues’ have low regeneration capacities, rendering them vulnerable to injury and osteoarthritis (OA), the most prevalent musculoskeletal disease that afflicts more than 27 million people in the US. Successful regeneration in articular cartilage and meniscus is challenged by the limited understanding of how these two tissues and their extracellular matrices (ECMs) initially form. This thesis queries the early molecular events that govern the assembly of the initial primitive matrix in the embryonic cartilage and meniscus. Using wild-type embryonic murine knee joint as the model system, we found that the primitive matrix of cartilage and meniscus initiates with a pericellular matrix (PCM)-like template that matures into the specialized PCM and bulk ECM regions. In this early phase, articular cartilage and meniscus developed molecular characteristics signifying distinct matrix templating during initial formation. Also, from embryonic to neonatal development, the micromodulus of cartilage and meniscus matrices stiffen exponentially, with a daily modulus increase rate of ≈ 36% and ≈ 20%, respectively. These results together highlighted the rapid and distinct development traits of the primitive matrices for these two tissues, providing the foundation for determining the molecular mechanisms governing the early matrix templating, and for developing novel tissue engineering strategies to recapitulate the native ECM formation.

Building on these findings, we examined the role of type V collagen, a minor fibril-forming collagen, in regulating the initial matrix templating of both tissues. Although collagen V is usually considered as a co-initiator of collagen I fibrillogenesis, we found pronounced expression in both collagen I-based meniscus and collagen II-rich articular cartilage at embryonic stage. In the joint-specific Col5a1 knockout model (Col5a1f/f/Gdf5Cre, or Col5a1cKO), we found substantial matrix defects in newborn tissues, including reduced meniscus size, flattened cartilage, thickened collagen fibrils and reduced tissue modulus.

Despite these changes, single-cell RNA-sequencing did not yield clear phenotype in cell phenotype or signaling, supporting a direct role of collagen V in initial matrix templating. Such impaired matrix templating results in disrupted postnatal growth. In 1-month-old Col5a1cKO joints, we found decreased meniscus size, an absent meniscus inner zone with much reduced Col2a1 expression, and reduced proteoglycan content in cartilage. These changes further progressed to marked signs of OA-like cartilage and meniscus degeneration by 3 months of age, as well as aberrant formation of osteophytes and subchondral bone remodeling, which are signs of advanced OA, by 8 months of age. Together, these results show that collagen V is an essential constituent of the initial matrix templating of both articular cartilage and meniscus. Targeting molecular activities of collagen V could hold the potential of improving the regeneration of these two structurally distinct tissues to treat OA and joint injury.

BIOMED Master's Thesis Defense

Title:
Developing Mutant KRAS Targeted Vaccines for Pancreatic Cancer Interception

Speaker:
Ben Barrett, Master's Candidate
School of Biomedical Engineering, Science and Health Systems
Drexel University
 
Advisors:
Neeha Zaidi, MD
Assistant Professor of Oncology
Sidney Kimmel Comprehensive Cancer Center
Johns Hopkins Medicine
 
Adrian Shieh, PhD
Teaching Professor
School of Biomedical Engineering, Science and Health Systems
Drexel University

Details:
Pancreatic Ductal Adenocarcinoma (PDAC) remains one of the most lethal cancers to date, with a 5-year survival rate of only 12%. The poor survival rate can largely be attributed to an immunosuppressive tumor microenvironment (TME) that is largely devoid of anti-tumor T cells. Approximately 90% of PDACs are driven by mutations in KRAS (mKRAS), with the most common being KRASG12D (~40%). mKRAS serves as an ideal set of candidates for targeting due to their role as a driver mutation and high specificity to precancerous lesions. These shared driver mutations, along with the near decade-long timeframe from pre-cancer to PDAC development, allows for a window-of-opportunity to develop vaccines to activate T cells before cancer develops and immunosuppression evolves. Several considerations are key to vaccine performance including the platform and immunomodulatory adjuvants admixed with neoantigens. Within this thesis, we explore two promising vaccine platforms: a pooled neoantigen peptide vaccine and a bicistronic mRNA vaccine. Each platform contains distinct benefits with peptide vaccines consistently demonstrating immune responses to neoantigens, while mRNA offers the possibility of directly encoding immunomodulatory adjuvants often required to be admixed with peptide vaccines for acceptable responses. The stimulator of interferon genes (STING) pathway offers an increasingly attractive target for optimizing T cell response due to the upregulation of type I interferons.

We thus designed and verified the in vitro functionality of an mRNA vaccine encoding both a KRASG12D vaccine and immunomodulatory adjuvant STINGV155M, a constitutively active STING (caSTING) protein, on the same vector mediated by an EMCV IRES linker. We set 3 requirements that were paramount to design success: 1) Antigenic translation (KRASG12D), 2) Adjuvant Translation (STINGV155M), and 3) Constitutive STINGV155M Activation. Two constructs were tested that alternated the ordering of the cistrons, and thus modified the nature of translation. It was determined that while both encoded proteins were translated from the same vector using Western Blot, there was insufficient caSTING functionality when caSTING was subject to cap-independent translation as measured by a STING reporter cell line. However, STING signaling was rescued when the caSTING was translated via cap-dependent initiation, indicating that bicistronic translation requires improvement before it can be deemed that single construct met all requirements.

In parallel, we characterized the immunogenicity of a pool of mutant KRAS synthetic long peptides (SLPs) (G12C, G12V, G12R, G12A, G12D and G13D) via anti-IFNγ ELISpot and flow cytometry T cell phenotyping in C57BL/6 mice. We demonstrated robust antigen-specific responses when the SLPs were admixed with a STING adjuvant. We also found an increase in mKRAS-specific T cell responses when increasing the number of doses in the vaccination protocol. Interestingly, we found cross-reactive T cell responses in both CD4+ and CD8+ T cells, dependent on the epitopes included in the peptide mixture.

Finally, we characterized the pre-malignant and tumor microenvironment (TME) of an inducible, conditional mouse model of PDAC, commercially available from the Jackson Laboratory, Pdx1-CreERTg/Tg;Trp53fl/fl;KrasG12D/+. This mouse model expands on the Cre-Lox recombination of the traditional KPC mouse model, through the association of an estrogen receptor with the Cre protein. Thus, the estrogen-analog, Tamoxifen, is required for Cre to enter the nucleus and carry out recombination, allowing for the induction of the KRASG12D-driven tumorigenesis. We developed a method of inducing mKRAS, isolating and maintaining tissue, and studying tissue histology. We confirmed the expression of KRASG12D localized to precancerous lesions (PanINs) and PDAC tumors through in situ hybridization (ISH). We further observed a migration of CD3+ and CD68+ cells to sites of PanIN and PDAC via immunohistochemistry (IHC) staining, specifically noting a clear integration of immune cells within PanIN lesions and a clear restriction of CD3+ cells to the border of tumors. Based on the data collected, the tiKPC model serves an ideal candidate for future immunotherapy interception studies using the vaccine formulations outlined above. Throughout the course of this thesis, we demonstrated the potential for mKRAS-targeted vaccine platforms with the intention of intercepting pancreatic cancer, as well as the ideal model to characterize the efficacy of an interception strategy.

Join OUTLaw for a finals study session during the reading period where we can all chill, breathe, and work together to survive this stressful time. Coffee and hot teas will be available, as well as breakfast pastries and fresh fruit.