Hello, my name is Montserrat Lara, I'm going to be showing the results of my project entitled Cerebral Spinal Fluid Responsive Factor Serpenate 3 Promotes Proliferation, Migration, and Invasion of Glioblastoma. Glioblastoma is the most common and aggressive form of primary brain cancer in adults. It accounts for 54% of new gliomas and 45% of primary malignant tumors. The standard of care includes surgery, chemotherapy, and radiotherapy, however, the survival expectancy is only 14 months and the recurrence rate is close to 100%. Tumor location is also a contributing factor for the dismissal prognosis of the disease. Periventricular GBM exhibits lower survival rate as well as higher capacity to migrate to distant sites from the primary tumor. The main reason for the worst survival in GBM periventricular tumors is not known, however, it has been suggested that the interaction between neurogenic niches and GBM tumors can be a contributing factor. In this project specifically, we were interested in studying the chemical codes immersed in the cerebrospinal fluid that can be regulating the GBM cell's migration. First, we evaluated the effect of cancer CSF, non-cancer CSF, and control condition in migration and proliferation of the GBM cells. We saw higher migration and proliferation rate upon cancer CSF stimulation. We also evaluated the transcriptome changes in cancer cells upon cancer CSF stimulation. As you can see here, we got a list of genes that were over-regulated upon this conditioning and we found genes such as aldoC, CD44, osteopointin, and serpenatry with higher expression in the cells. These genes have been related previously with cancer. We also studied the protein composition itself of the samples through mass spectrometry and then we identified among the cancer CSF groups 134 proteins that were shared in common. Interestingly, in one of the main clusters of proteins, we identified again serpenatry. What is serpenatry? It's a serine protease inhibitor that inhibits scatopsin G and muscle chemosis. It's synthesized by the liver, astrocytes, and endometrial cells and is increased in inflammatory processes, neurodegenerative diseases, and cancer. In cancer such as endometrial, melanomic, gastric cancer, and glioma, it's overexpressed. In glioma, serpenatry expression is correlated with glioma grade, a higher grade, higher expression of serpen. Serpenatry also correlates with survival. A higher expression of the protein lowered the survival. It doesn't differ the expression of the protein among molecular subtypes of GVM, and again, we confirmed here that the expression of serpen correlates with glioma grade. We identified that cancer GVM samples had higher expression of the protein and at the mRNA level of serpenatry when compared with the non-cancer samples. We also detected the expression of serpenatry among different V-ticks from glioma patients. And we identified higher protein level concentration in different high-grade CSF samples when compared with non-cancer CSF samples. We successfully knocked down serpenatry in three V-ticks, and as you can see here, the knockdown of the protein impairs migration and invasion capabilities of the cancer cells. We also saw a decreased clonal capacity in the knockdown cells as well as a decreased synthesis phase on the cell cycle and colony formation ability. Interestingly, when we differentiate the cells, we detected lower levels of serpenatry at the mRNA level as well as several stem cell-related markers. Through flow cytometry, we identified the positive and negative CD133 population of V-ticks, and we saw a higher mRNA levels of serpenatry in the positive population. And after silencing serpenatry, we detected decreased CD133 expression in the cells. Through LMR-BLUE analysis, we saw that serpenatry knockdown decreases viability in the cancer cells. In vivo, we transplanted the cells, the controls, and the knockdown groups, and we saw higher survival rate in the knockdown group as well as decreased CHI-67 proliferation index after silencing serpenatry. We also detected lower invasion capacity, meaning lower capacity to migrate through the corpus callosum to the contralateral hemisphere in the knockdown groups for serpenatry. I want to thank all the members in my lab, my PI, and I want to thank also the patients who made this project possible by donating their samples. Thank you very much. This is the funding that supported the project. Thank you.

Montserrat Lara

Cerebral Spinal Fluid Responsive Factor Serpenatry 3

Glioblastoma

Chemical codes

CSF