Quantifying Neuronal Damage via Immunofluorescence Staining in Chemotherapy Induced Peripheral Neuropathy (CIPN)
Abstract
Multiple myeloma is the second most common hematological cancer of plasma cells and Bortezomib (BTZ), a proteasome inhibitor, is commonly used to treat this condition. BTZ causes debilitating Chemotherapy Induced Peripheral Neuropathy (CIPN) that results in the dose limitation or discontinuation of chemotherapy. Our study focuses on determining BTZ induced neuronal damage using PC-12 cells, derived from pheochromocytoma found in rats. PC-12 cells qualify as a viable in vitro neuronal model due to their ability to differentiate into neurons with the addition of nerve growth factor (NGF). The neuronal markers including TUBB3 (β-Tubulin), GFAP (Glial Fibrillary Acidic Protein), MBP (Myelin Basic Protein) and NeuN (Neuronal nuclei) were used in immunochemical staining to confirm differentiated PC-12 cells’ neuronal lineage. High expression of the biomarkers (CREB and NSE) was observed using protein quantification techniques (Western Blot), confirming significant neuronal damage in the BTZ-treated PC-12 cells. Additionally, immunofluorescence staining was used to measure the neuronal damage induced by BTZ. Immunofluorescent staining of the damaged neurons under a fluorescence microscope, allowed us to accurately characterize and quantify the changes in neurite outgrowth, in response to chemotherapeutic agents as well as potential neuroprotective treatments. Furthermore, our results will be used to measure the effectiveness of neuroprotective treatments such as vitamin B12, gamma linolenic acid (GLA), and neuronal nitric oxide synthase (nNOS) inhibitor. (This project was supported by the President’s Faculty Research and Development Grant from NSU and the generous financial support from The Royal Dames of Cancer Research Inc., Ft. Lauderdale, Florida).
Faculty Sponsors
Dr. Paramjot Kaur, Dr. Theodore Mathuram, Dr. Thiagarajan Venkatesan, Dr. Mir Saleem, Dr. Appu Rathinavelu
Project Type
Event
Location
Alvin Shermany Library
Start Date
4-5-2019 1:00 PM
End Date
4-5-2019 5:00 PM
Quantifying Neuronal Damage via Immunofluorescence Staining in Chemotherapy Induced Peripheral Neuropathy (CIPN)
Alvin Shermany Library
Multiple myeloma is the second most common hematological cancer of plasma cells and Bortezomib (BTZ), a proteasome inhibitor, is commonly used to treat this condition. BTZ causes debilitating Chemotherapy Induced Peripheral Neuropathy (CIPN) that results in the dose limitation or discontinuation of chemotherapy. Our study focuses on determining BTZ induced neuronal damage using PC-12 cells, derived from pheochromocytoma found in rats. PC-12 cells qualify as a viable in vitro neuronal model due to their ability to differentiate into neurons with the addition of nerve growth factor (NGF). The neuronal markers including TUBB3 (β-Tubulin), GFAP (Glial Fibrillary Acidic Protein), MBP (Myelin Basic Protein) and NeuN (Neuronal nuclei) were used in immunochemical staining to confirm differentiated PC-12 cells’ neuronal lineage. High expression of the biomarkers (CREB and NSE) was observed using protein quantification techniques (Western Blot), confirming significant neuronal damage in the BTZ-treated PC-12 cells. Additionally, immunofluorescence staining was used to measure the neuronal damage induced by BTZ. Immunofluorescent staining of the damaged neurons under a fluorescence microscope, allowed us to accurately characterize and quantify the changes in neurite outgrowth, in response to chemotherapeutic agents as well as potential neuroprotective treatments. Furthermore, our results will be used to measure the effectiveness of neuroprotective treatments such as vitamin B12, gamma linolenic acid (GLA), and neuronal nitric oxide synthase (nNOS) inhibitor. (This project was supported by the President’s Faculty Research and Development Grant from NSU and the generous financial support from The Royal Dames of Cancer Research Inc., Ft. Lauderdale, Florida).
