1. Tumor-stromal Interactions in Metastatic Brain Cancer Kathryn Suttling Mentor: Victor Blanco PI: Xiaoyang Qi, PhD Lab Department: Internal Medicine/ Hematology and Oncology Vontz Center for Molecular Studies

2. Our Model Nude Mouse model Skin Cancer Cells 3-4 Weeks Brain Slides Immunofluorescence Dissection and Sectioning of the mouse brain Intracarotid Artery Injection CD11b 1 st AB (Rabbit) against Ki-67 1 ry AB (Rat) against CD11b 2 ry AB-TRITC (anti-mouse) 2 ry AB-FITC (anti-rabbit) Ki-67

3. Histology of Experimental Melanoma Brain Metastases (H&E) H & E: Routine histopathological characterization to identify tumor cells. 4X 10X 40X Normal- 10X

4. Stromal Cells in Melanoma Brain Metastases Immunofluorescence DAPI: Marks nuclei (DNA). Dense areas of cells compared to different regions are generally tumorous. GFAP: High expression indicates reactive astrocytes. Nestin: Stem cell marker- Co-option of host cells. 40X DAPI/Nestin Merge DAPI GFAP Normal 4X 40X 4X

5. Immune Cells in Melanoma Brain Metastases CD11b: Labels macrophages/microglia (Tumor Associated Macrophages) Ki-67 : Labels proliferating cells Versican: Chondroitin sulfate; matrix component; possible immune activator Ki-67 CD11b DAPI Versican DAPI Merge 4X Merge 4X

6. Summary  Valuable model to study the progression and interaction of brain metastases of melanoma and stromal and immune cells.  Through immunofluorescence techniques, we identified astrogliosis, tumor’s co-option of host cells and immune cell involvement in melanocytic brain metastases.  Next steps are to figure out how to make tumor cells to uptake SapC-DOPS (anticancer liposomes). Right now there is targeting, but only around the tumors.  In the future, we will be looking for the potential therapeutic applications of SapC-DOPS

7. My Experience  What have I learned?  Research process:  Trial and error  Experiments must be validated  Research takes time  About Myself:  In order to be good at something, work and time must be put into it.  Why does it matter ?  To be good in something, you need to put in the work  Passion

8. My Experience  What now?  Academic: Use basic lab techniques in future science classes. Gain an understanding of what is happening when an experiment is performed.  Professional: As I look towards graduate school, the option of engaging research will be available to me if I so choose.  Personal: This has taking me one step closer to figuring out what direction I want to take in life.