Skip to main content
2024 BMES Annual Meeting Main banner
Icon Legend
This session is not in your schedule.
This session is in your schedule. Click again to remove it.

High School Student with a Traditionally Underrepresented Background

HS Poster

Poster 16 - Analysis of Immune Involvement in Pancreatic Ductal Adenocarcinoma in Genetically Engineered Mice

Saturday, October 26, 2024
10:00 AM - 12:00 PM EST
Location: Exhibit Hall E, F & G

Introduction: Pancreatic cancer stands out among cancers for being one of the most deadly post detection. This is partially due to current treatment methods being inadequate or ineffective, with chemotherapy unable to penetrate the tumor and surgical resection / radiotherapy bringing the risk of damage to other vital organs. As such, a novel therapeutic approach lacking the side effects and risks of current therapies is necessary to improve patient outcomes. Immunotherapies are treatment methods that have been used to great effect on normal diseases and cancers alike. They do not entail many of the side effects that make current treatments unappealing, and can prove very effective. So, this study aims to evaluate the foundational viability of immunotherapies to treat pancreatic cancer, utilizing genetically engineered mouse models, and both qualitative and quantitative methods of analysis.

Materials and

Methods: KrasLSL-G12D;p53LoxP;Pdx1-CreER (KPC) Mouse (genetically engineering to develop humanlike pancreatic ductal adenocarcinoma (PDA)) tissue samples:
- Spleen for positive immune presence control, and pancreas as organ of interest for testing immune presence
- 2 early stage PDA-afflicted mice dissected to be used in tissue digestion then flow cytometry
- 3 late stage PDA-afflicted mice pre-prepared in slides (prepared some time ago by graduate researchers) to be used in immunofluorescent staining and hematoxylin and eosin staining
Immunofluorescence (IF) / hematoxylin and eosin (H&E) staining:
- In IF, late stage mouse models were prepared in slides then bound with B cell and dendritic cell primary antibodies to qualitatively measure amounts of those cells. Secondary antibodies allowed fluorescence of target immune cells subsets under laser excitation, enabling fluorescent microscopy for imaging
- In H&E staining, late stage mouse models were prepared in slides then dipped through an H&R staining array to allow structural analysis of PDA tumor sites under bright field microscopy
Flow cytometry (using BSD LSRFortessa X-20):
- The 2 early stage KPC mice were dissected, and their spleens and pancreases placed into an enzyme solution for tissue digestion (collagenase and hyaluronidase).
- Solutions were incubated then washed, centrifuged, and placed into single cell suspension
- These suspensions were incubated with conjugated antibodies to ensure fluorescence during flow cytometry
- Washing and fixing steps with phosphate buffered saline and paraformaldehyde to prep suspension for flow cytometry
- Suspensions analyzed in 3 channel flow cytometry using flow compensation beads for single stain controls

Results, Conclusions, and Discussions: According to the data from flow cytometry procedure, it indicates lessened counts of both B cells and T cells in pancreas when compared to spleen, but significantly increased counts of dendritic cells in pancreas when compared to spleen.

Relatively larger (than that of other immune cells) presence of dendritic cells in both early and late stage PDA (shown in both flow cytometry (Fig. 5) and IF data(Fig. 4)), indicating the immune system identified tumor as foreign object and attempted to present antigen to initiate immune response targeted at cancer cells
IF staining of pancreas (Fig.4B) revealed lessened immune presence as compared to spleen (Fig. 4A), suggesting immune exclusion as tumor developed

While the presence of dendritic cells in the pancreas during early stage cancer is promising, the relative lack of B cells and T cells signals a need for future therapies to enable further immune infiltration into pancreases and pancreatic tumors via modification of immune cells.

Acknowledgements (Optional): We would like to sincerely thank our mentors, Brianna Brennecke and Noah Sinclair, as well as Paolo Provenzano and his lab for their invaluable guidance and support throughout our project.

References (Optional): 1) Luu, A. M et al., (2021, April). Late recurrences of pancreatic cancer in patients with long-term survival after pancreaticoduodenectomy. Journal of gastrointestinal oncology.
2) Grove, E. (2023, October 2). Mayo Clinic's AI innovation inspires hope in early detection of pancreatic cancer. Mayo Clinic News Network.
3) Pancreatic cancer - Diagnosis and treatment. (2024, May 4). Mayo Clinic.
4) Howard, C. J., Charleston, B., Stephens, S. A., Sopp, P., & Hope, J. C. (2004). The role of dendritic cells in shaping the immune response. Animal health research reviews, 5(2), 191–195.
5) Janeway CA Jr, Travers P, Walport M, et al. Immunobiology: The Immune System in Health and Disease. 5th edition. New York: Garland Science; 2001. Chapter 8, T Cell-Mediated Immunity.
6) Althwaiqeb, S. A., & Bordoni, B. (2023, May 29). Histology, B Cell Lymphocyte - StatPearls. NCBI.
7) Figures created with BioRender.com