School of Medicine and Health Sciences Poster Presentations

Title

Expression of HDAC10 and HDAC11 in lymphoma and myeloma and potential therapeutic implications.

Document Type

Poster

Abstract Category

Cancer/Oncology

Keywords

lymphoma, myeloma, HDAC

Publication Date

Spring 5-1-2019

Abstract

New therapies are desperately needed to improve patient response and prevent patient relapse in lymphoma and myeloma. Histone Deacteylases (HDACs) are potential therapeutic targets that could greatly improve cancer treatment. Aberrant expression of HDAC10 and HDAC11 have been noted in several hematologic malignancies. Specifically, the protective role of HDAC11 in myeloma is well-known, and may also exist in lymphoma. Thus, we hypothesized that both HDAC10 and HDAC11 play an important role in hematologic malignancies. The project involved screening T and B cell lymphoma lines and myeloma cell lines for HDAC10 and HDAC11 RNA expression, and using a gene expression profile of various lymphomas to compare HDAC10 and HDAC11 expression with a normal control. Methods: Fourteen human cancer cell lines were cultured in RPMI-1640 media with 10-20% FBS and 1% penicillin/streptomycin according to American Type Culture Collection (ATCC) guidelines.. The diffuse large B cell lymphoma (DLBCL) cell lines SU-DHL-4, SU-DHL-6, HBL-1, OCI-LY-3, OCI-LY-19, and U2932 were used. The multiple myeloma cell lines Toledo and U266 were used. Finally, the T cell lymphoma lines used were HH, MYLA, and SR786. RNA was harvested from cells using the Maxwell 16 LEV simplyRNA Cell Kit.. cDNA synthesis was performed from 2000 ng RNA samples using the iScript cDNA synthesis kit. Finally, qRT-PCR was done using a Thermo Fisher thermocycler, quiagen HDAC10 (QT00007252) and HDAC11 (QT01674617) primers, and SYBR green master mix from Thermo Fisher. A one-way ANOVA test was performed to determine if any differences in expression existed between the three cell types (B cell lymphomas, T cell lymphomas, and multiple myeloma), followed by an unpaired T test to directly compare two groups together. A gene expression profile dataset from the Spanish National Cancer Research Center (CNIO) was used for HDAC expression analysis. All patients gave informed consent, and all information was de-identified. The dataset included the following samples: 11 reactive lymphoid tissue(controls), 24 MCL, 25 follicular lymphoma, 16 CLL, 11 nodal marginal zone lymphoma, 15 mucosal-associated lymphoid tissue lymphoma, 19 splenic marginal zone lymphoma, 25 DLBCL, 24 T cell lymphoma samples (TCL), 6 lymphaplasmacytoid lymphoma samples, and 4 Burkitt's lymphoma. Results The PCR data showed no significant difference between HDAC11 expression between the different types of lymphoma and multiple myeloma. However, the data did show a statistically significant difference between HDAC10 expression between T cell lymphomas and multiple myeloma, with MM expressing slightly higher levels of HDAC10 than T cell lymphoma lines (p value=.0153). The gene expression profiling data showed a significantly higher expression of HDAC10 in MCL samples from all other cell types. HDAC10 expression was significantly decreased in

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Presented at Research Days 2019.

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Expression of HDAC10 and HDAC11 in lymphoma and myeloma and potential therapeutic implications.

New therapies are desperately needed to improve patient response and prevent patient relapse in lymphoma and myeloma. Histone Deacteylases (HDACs) are potential therapeutic targets that could greatly improve cancer treatment. Aberrant expression of HDAC10 and HDAC11 have been noted in several hematologic malignancies. Specifically, the protective role of HDAC11 in myeloma is well-known, and may also exist in lymphoma. Thus, we hypothesized that both HDAC10 and HDAC11 play an important role in hematologic malignancies. The project involved screening T and B cell lymphoma lines and myeloma cell lines for HDAC10 and HDAC11 RNA expression, and using a gene expression profile of various lymphomas to compare HDAC10 and HDAC11 expression with a normal control. Methods: Fourteen human cancer cell lines were cultured in RPMI-1640 media with 10-20% FBS and 1% penicillin/streptomycin according to American Type Culture Collection (ATCC) guidelines.. The diffuse large B cell lymphoma (DLBCL) cell lines SU-DHL-4, SU-DHL-6, HBL-1, OCI-LY-3, OCI-LY-19, and U2932 were used. The multiple myeloma cell lines Toledo and U266 were used. Finally, the T cell lymphoma lines used were HH, MYLA, and SR786. RNA was harvested from cells using the Maxwell 16 LEV simplyRNA Cell Kit.. cDNA synthesis was performed from 2000 ng RNA samples using the iScript cDNA synthesis kit. Finally, qRT-PCR was done using a Thermo Fisher thermocycler, quiagen HDAC10 (QT00007252) and HDAC11 (QT01674617) primers, and SYBR green master mix from Thermo Fisher. A one-way ANOVA test was performed to determine if any differences in expression existed between the three cell types (B cell lymphomas, T cell lymphomas, and multiple myeloma), followed by an unpaired T test to directly compare two groups together. A gene expression profile dataset from the Spanish National Cancer Research Center (CNIO) was used for HDAC expression analysis. All patients gave informed consent, and all information was de-identified. The dataset included the following samples: 11 reactive lymphoid tissue(controls), 24 MCL, 25 follicular lymphoma, 16 CLL, 11 nodal marginal zone lymphoma, 15 mucosal-associated lymphoid tissue lymphoma, 19 splenic marginal zone lymphoma, 25 DLBCL, 24 T cell lymphoma samples (TCL), 6 lymphaplasmacytoid lymphoma samples, and 4 Burkitt's lymphoma. Results The PCR data showed no significant difference between HDAC11 expression between the different types of lymphoma and multiple myeloma. However, the data did show a statistically significant difference between HDAC10 expression between T cell lymphomas and multiple myeloma, with MM expressing slightly higher levels of HDAC10 than T cell lymphoma lines (p value=.0153). The gene expression profiling data showed a significantly higher expression of HDAC10 in MCL samples from all other cell types. HDAC10 expression was significantly decreased in