1.6 Hodgkin’s Lymphoma
Previous in silico research in our laboratory found that H2A.B is highly expressed in one particular cancer type, Hodgkin’s Lymphoma. Hodgkin’s Lymphoma (HL) is a type of B cell cancer that affects both men and women of various ages. HL can be divided into two classes: classical HL (cHL) and Nodular Lymphocyte Predominant HL (NLPHL) according to the WHO classification 2008 (Swerdlow et al., 2008). The classical HL accounts for 95% of all HL cases, and it has been further divided into 4 subtypes: Nodular sclerosis cHL, which is the most common phenotype (75% of cHL cases); Mixed cellularity cHL, the second most common phenotype (15-25% of cHL cases); Lymphocyte Rich cHL and Lymphocyte Depleted cHL, which comprise 6% and …show more content…
CD15 and CD30 are the currently using marker that are expressed on most of the HRS cells. However, CD15 and CD 30 cannot be used for non-classical HL (Lymphocyte predominant HL) diagnosis because the pathogenic L&H cells are negative for both CD15 and CD30 (Zhao, 2008). Another limitation is that CD15 and CD30 were also observed to be positive for few normal cell types and cancer cells. For example, both CD15 and CD30 are shown positive to activated B cells and T cells, as well as a group of other types lymphoma (Ito et al., 1994, Lim et al., 2002 and Barry et al., 2003). Current treatments of HL are multi-agent chemotherapy and radiation. These treatments can generally achieve a 65-90% cure rate (Diehl, thomas and Re, 2004). However, these treatments normally cause severe side effects. In order to reduce the toxicity, recent research has focused on developing antibody drug conjugate to target HRS cells (Foyil and Bartlett, 2010). A monoclonal antibody against CD30 conjugated with drugs has been tested with patients and has showed promising result. H2A.B is a protein, which is specifically expressed in HL pathogenic cells. Understanding of H2A.B functions in HL cells may contribute to our overall understanding of HL and the development of potential cures in the …show more content…
Characterize the expression of H2A.B in HL patients. To achieve this, the expression of H2A.B will be visualized by immunohistochemistry staining in 81 Hodgkin’s Lymphoma patients.
2. Characterize the HL cell lines as a new model for studying H2A.B and characterize the overall expression of H2A.B in 3 HL cell lines. This will be achieved by confirming the H2A.B coding gene integrity through sequencing and determining the H2A.B expression level by qPCR and Western Blot.
3. Generate the H2A.B knockdown cells to uncover the function of H2A.B. H2A.B expression will be down-regulated by transducing HL cell lines with Lentiviral shRNA constructs. The impacts of H2A.B down-regulation will be assessed by observing the differences in chromatin compaction and gene expression when comparing knockdown cells to the wild type cells.
1.8 The significance of this
Previous in silico research in our laboratory found that H2A.B is highly expressed in one particular cancer type, Hodgkin’s Lymphoma. Hodgkin’s Lymphoma (HL) is a type of B cell cancer that affects both men and women of various ages. HL can be divided into two classes: classical HL (cHL) and Nodular Lymphocyte Predominant HL (NLPHL) according to the WHO classification 2008 (Swerdlow et al., 2008). The classical HL accounts for 95% of all HL cases, and it has been further divided into 4 subtypes: Nodular sclerosis cHL, which is the most common phenotype (75% of cHL cases); Mixed cellularity cHL, the second most common phenotype (15-25% of cHL cases); Lymphocyte Rich cHL and Lymphocyte Depleted cHL, which comprise 6% and …show more content…
CD15 and CD30 are the currently using marker that are expressed on most of the HRS cells. However, CD15 and CD 30 cannot be used for non-classical HL (Lymphocyte predominant HL) diagnosis because the pathogenic L&H cells are negative for both CD15 and CD30 (Zhao, 2008). Another limitation is that CD15 and CD30 were also observed to be positive for few normal cell types and cancer cells. For example, both CD15 and CD30 are shown positive to activated B cells and T cells, as well as a group of other types lymphoma (Ito et al., 1994, Lim et al., 2002 and Barry et al., 2003). Current treatments of HL are multi-agent chemotherapy and radiation. These treatments can generally achieve a 65-90% cure rate (Diehl, thomas and Re, 2004). However, these treatments normally cause severe side effects. In order to reduce the toxicity, recent research has focused on developing antibody drug conjugate to target HRS cells (Foyil and Bartlett, 2010). A monoclonal antibody against CD30 conjugated with drugs has been tested with patients and has showed promising result. H2A.B is a protein, which is specifically expressed in HL pathogenic cells. Understanding of H2A.B functions in HL cells may contribute to our overall understanding of HL and the development of potential cures in the …show more content…
Characterize the expression of H2A.B in HL patients. To achieve this, the expression of H2A.B will be visualized by immunohistochemistry staining in 81 Hodgkin’s Lymphoma patients.
2. Characterize the HL cell lines as a new model for studying H2A.B and characterize the overall expression of H2A.B in 3 HL cell lines. This will be achieved by confirming the H2A.B coding gene integrity through sequencing and determining the H2A.B expression level by qPCR and Western Blot.
3. Generate the H2A.B knockdown cells to uncover the function of H2A.B. H2A.B expression will be down-regulated by transducing HL cell lines with Lentiviral shRNA constructs. The impacts of H2A.B down-regulation will be assessed by observing the differences in chromatin compaction and gene expression when comparing knockdown cells to the wild type cells.
1.8 The significance of this