Epigenetics and Cancer

by Prof. Dr. Motawa E. El Houseini

It is a series of functional unit of genes, each of which contains the information for a single function. The genes were expressed via mRNA and translated to proteins which carry out the biological activities of the cells. The gene is structural unit of the genome which contains the following regions: - Promoter region. -Enhancer. -Silencer sequence.

Genome:

 Definition: Epigenetics represents the science for the studying heritable changes of DNA, not involving changes in DNA sequence that regulate gene expression. There are at least two forms of information in the genome of the cell. A- Genetic information B- Epigenetic information

 The former provides the building block for the manufacture of all Proteins needed for the cell functional activity. The latter provides additional instruction on how, when and where these information should be used.

Biochemical reactions which are operating in Epigenetics 1- DNA methylation and demethylation 2- Histone acetylation and deacetylation. 3- Histone methylation and demethylation 4- Phosphorylation and dephosphorylation of histones and non-histone proteins

Biochemical reactions which are operating in Epigenetics 1- DNA methylation and demethylation 2- Histone acetylation and deacetylation. 3- Histone methylation and demethylation 4- Phosphorylation and dephosphorylation of histones and non-histone proteins

First DNA methylation

It is the covalent addition of methyl group to 5th Position of cystosine with in CPG dinucleotides which are frequently located in the promoter region of genes. It is a complex process catalyzed by DNA methyl transferase. The addition of the methyl group from the universal methyl Donor s-adenosyl L -methionine.

Epigonome and acetylation

Epigenetic changes of the genome include: A- DNA methylation as mentioned before. B- Chromatin remodeling by histone modification by acetylation. The acetylation of histones (H) mainly H3 and H4 is done at A.A lysine 9 & 14 and lysine 5, 8, 12, 16 respectively by enzyme systems histone acetyltransferases.

The latter transfer the acetyl co-enzyme A to the lysine residue which leads to neutralizing the positive charge. Histone acetylation loosens chromatin packaging and correlates with transcriptional activation. Whereas, histone deacetylases remove the acetyl groups re-establishing the positive charge in the histones which are associated with repression of transcription.

Histone methylation
The process is carried out by an enzyme histone methyl transferase which directs site-specific methylation of amino-acid residues such as lys. 4&9 in the tail of the histone H3.

- Methylation of lysine 9 in histone H3 directs the binding of noncoding RNA, histone deacetylase to control chromatin structure and gene expression.

 Neoplastic Transformation
It is a complex multi-event and multi-stage Process. The process can be divided into two requisite sequences 1- Neoplastic conversion 2- Neoplastic development The stages of the process can be shown as Follow:

Neoplastic Conversion
Chemical Carcinogen -DNA Reactive -Epigenetic effect -DNA methylation -Histone deacetylation Progression Promotion

Neoplastic cell

DNA alteration
Genetic and epigenetic Levels

Neoplastic Development

Genes promoters hypermethylation in human cancers

- The list of genes that are found to be inactivated by DNA methylation events is growing rapidly and includes genes involved in the following : A- Signal transduction cascade pathways. B- Cell cycle regulation. C-Angiogenesis. D-Apoptosis. E- DNA repair.

Recent Cancer methylation studies predict that hundred (100) of CPG islands could be methylated in a tumor cell. However, it is clear that both the genome-wide methylation studies and candidate gene approaches that each tumor type may have its own set of cancer cell type specific genes that are more susceptible to methylation. Thus each cancer type may have the potential to be typed or classified according to methylation profile.

P 15/P16 Methylation in cancer - Aberrant methylation of cycline dependent kinase inhibitor P16 INK 4a has been frequently detected in many human cancers. - P15INK4b gene adjacent to P16 on chromosome q p21, is also commonly hypermethylated in human neoplasm, (Hematological malignancies and head and neck sequamous cell carcinoma).

- The differential levels of methylated P16 and P15 in plasma might be potential useful markers in screening high risk population for an early detection of head and neck squamous cell carcinoma.

 In colorectal cancer: - Progressive methylation of DNA and subsequent silencing of a subset of genes occurs in normal tissues along side age and time dependent events which predispose these normal cells to neoplastic transformation.

 Cytosine methylation of CPG loci underlies many of the genetic events in colorectal carcinogenesis which can be discovered in very early stage. The inactivation of adenomatous polyposis coli (Apc): The gene is believed to initiate colorectal carcinogenesis in 80% of the cases therefore, it has potential as sensitive marker of on early stage of the disease.

In lung cancer
It has been shown that a panel of markers, for aberrant methylation that detects lung cancer at the early stages of development has been observed. This panel includes the following genes: -P 16 -APC -G-ST -E-cadhrin

- In breast cancer
BRCA gene is a breast cancer susceptibty gene, that is tumor suppressor gene responsible for both normal development and carcinogenesis in breast. BRCA1, reveals multi functional protein involved in DNA repair. Cell cycle regulation, transcription and apoptosis BRCA1 mutations may play a significant role in the tumor-genesis of familial breast cancer.

 Aberrant methylation of BRCA1 CPG island Promoter is associated with decreased BRCA1 mRNA in sporadic breast cancer cells.

 There is a strong relationship between BRCA1 promoter hyper-methylation and the existence of LOH (loss of hetero-zygosity) at the BRCA locus. This finding suggested that one allele has been lost by deletion and the other is inactivated by aberrant methylation. Both events simultaneously leading to the bi-allelic inactivation and complete lack of function of the BRCA1 gene.

 The frequent loss of BRCA1 protein In sporadic breast carcinoma could also result from epigenetic inactivation of both parental alleles. Reduced BRCA1 protein expression in tumors of patients with poor survival prognosis provides the evidence of this protein as a prognostic factor in patients with sporadic breast carcinoma.

 Targeting DNA methylation and histone acetylation. Decitabine drug (5 AZa-2 deoxycytidine) has been widely used as a demethylating agent in vitro and is used clinically in the treatment of acute leukemias and myelodysplasia. Therapetuc strategy was preceded in animal model using anti-sense oligonuleotides against DNA methyl transferase. The studies showed an inhibition of tumor growth and re-expression of p16 in treated animals. Combination of histone deacetylase inhibitors (HDAC) e.g. Trichostatin and DNA methy-transferase inhibitor decitabine exerts a synergistic effect on the re-expression of hyper methylated silenced genes such as p15, TIMP and DNA mismatch repair genes.

Epigenome in relation to cancer drug resistance

Intrinsic and acquired drug resistance remain the most unpredictable factor affecting chemotherapy. DNA hypermethylation has been found to be associated with drug resistance acquired during cancer chemotherapy and therefore, re-expression of methylation-silenced genes resulted in increased sensitivity to existing chemotherapy.

 Example: Many chemotherapeutic agents kill cancer cells through induction of apoptosis accordingly; deregulation of any gene involved in the activation of the apoptotic process may be a major mechanism of chemo-resistance. Thus treatment of cancer cells with demethylating agent (decitobine) could lead to re-expression of caspase 8 and restore the sensitivity to the chemotherapy. This suggests that targeting the silencing mechanism of apoptosis related genes may increase the efficacy of various cancer chemotherapy.

CONCLUSION
Hyper methylation of CPG islands associated With tumor suppressor genes has been proposed to contribute to carcinogenesis. Epigenetic markers open new avenues for an early detection, diagnosis , prognosis as well as therapeutic targets in cancers.

 DNA demethylating agents and histone deacetylase inhibitors are underway to awake those silent tumor suppressor genes for a better treatment of patients suffering from cancers .

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