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

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

1st AB (Rabbit)

against Ki-67

1ry AB (Rat) against CD11b

2ry AB-TRITC (anti-mouse)

2ry AB-FITC (anti-rabbit)

Ki-67

Histology of Experimental Melanoma Brain Metastases (H&E)

H & E: Routine histopathological characterization to identify tumor cells.

4X

10X

10X

40X Normal- 10X

Stromal Cells in Melanoma Brain MetastasesImmunofluorescence

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

Immune Cells in Melanoma Brain MetastasesCD11b: Labels macrophages/microglia (Tumor Associated

Macrophages)Ki-67: Labels proliferating cellsVersican: Chondroitin sulfate; matrix component; possible immune activator

Ki-67 CD11bDAPI

VersicanDAPI Merge

4X Merge

4X

4X

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

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

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.