https://openworks.mdanderson.org/sumexp21/1221/thumbnail.jp

Al Atrash, Gheath

Dawson, Nicole

Gibson, Amber Leah

St. John, Lisa

Sukhumalchandra, Pariya

English

OpenWorks @ MD Anderson

Methods:• Standard cell culture procedure was done for NSCLC, OS, U937 (histiocytic leukemia) and T2 (multiple myeloma) cell lines.• Reverse transcriptase - polymerase chain reaction (RT-PCR) was performed to assess endogenous CG mRNA.• Cells were pulsed with a 10:1 ratio of PMNs to malignant cell for 4 hours.• Cells were stained with appropriate antibodies, and we then performed flow cytometry to determine CG uptake.• Cytotoxic Assay was performed to assess percent killing when cells were co-incubated with CG1 specific cytotoxic lymphocytes (CTLs).Conclusions:• Both OS and NSCLC show no endogenously express CG• OS and NSCLC can take up CG when pulsed with irradiated PMNs• The cytotoxicity assay was inconclusive, likely due to a high concentration of CTLs.• In the future, we will validate CG uptake in multiple OS and NSCLC cells lines and hope to demonstrate dose-dependent cytotoxicity of target cells.Novel Immunotherapy Targets Cathepsin G in Solid TumorsAuthors: N. Dawson1, A. Gibson2, P. Sukhumalchandra1, G. Al Atrash1, L. St. John1 1 Department of Hematopoietic Biology & Malignancy, The University of Texas MD Anderson Cancer Center, Houston, TX2 Department of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, TXIntroduction:• Cathepsin G (CG) is a serine protease found in Polymorphonuclear Neutrophils (PMNs).• Research suggests some leukemia tumor cells can take up CG and present its respective peptide (CG1) on their surface.• We hypothesize that CG can be taken up by Non-Small Cell Lung Cancer (NSCLC) cells and Osteosarcoma (OS) cells and be presented on their surface as novel immunotherapy targets.Figure 2: A. CG uptake H1650 Non-Small Cell Lung Cancer cells (pulsed at a ratio of 10:1). B. CG uptake in SaOS2 Osteosarcoma cells (pulsed at a ratio of 7:1)Figure 1: RT-PCR show no endogenous CG MRN in OS or NSCLC cell linesResults:• RT-PCR showed no endogenous mRNA expression of CG in NSCLC/OS cells lines (Figure 1).• Flow cytometry analysis in H1650 (NSCLC) showed CG uptake with a 1.25-fold increase in the CG-mean fluorescence intensity (CG-MFI) vs. non-pulsed cells (Figure 2).• Flow cytometry analysis in Saos2 (OS) showed CG uptake with a two-fold increase in the CG-MFI vs. non-pulsed cells (Figure 2).• Cytotoxicity assay showed non-specific killing of both pulsed and control (T2) control cells (Figure 3).Cathepsin GActinFigure 3: Cytotoxic Assay A. T2 control B. H1650 NSCLC

Novel Immunotherapy Targets Cathepsin G in Solid Tumors

https://openworks.mdanderson.org/cgi/viewcontent.cgi?article=1221&context=sumexp21

Novel Immunotherapy Targets Cathepsin G in Solid Tumors

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