Method for the determination of the DNA methylation level of a CPG position in identical cells within a tissue sample
AssigneeEpigenomics AG
InventorsJoern Lewin, Kurt Berlin
Life of this patent
Estimating the term…
Abstract
Aspects of the present invention relate to the determination of the DNA methylation level at one or more CpG position within cells of a defined type in a tissue sample. This methylation level is deduced from the total DNA methylation level of all cells of the sample and from the content of said cells of interest. In aspects of the invention, the cell content is determined by means of histopatholoy, staining methods, antibodies, expression analysis or DNA methylation analysis.
Claims32
Method for the determination of the DNA methylation level of one or more specific CpG positions of a subpopulation of cells within a tissue sample, comprising
- a) determining the cell content of said subpopulation of cells within the tissue sample in a quantitative or semi-quantitative manner,
- b) measuring the total DNA methylation level of said one or more specific CpG positions within the tissue sample,
- c) determining the DNA methylation level within the cells of said subpopulation of cells from the total DNA methylation level of said one or more specific CpG positions within the tissue sample and the cell content of said subpopulation of cells within the tissue sample.
Method for diagnosing a condition or disease, characterized by specific methylation levels of one or more methylation variable genomic DNA positions in a disease-associated cell of a tissue sample, comprising:
- a) obtaining a tissue sample comprising genomic DNA having one or more methylation variable positions in one or more regions thereof,
- b) determining the disease-associated cell content within the tissue sample in a quantitative or semi-quantitative manner,
- c) measuring the total DNA methylation level of one or more methylation variable genomic DNA positions of the tissue sample,
- d) determining the DNA methylation level of one or more methylation variable genomic DNA positions within the disease-associated cells from the total DNA methylation level and the disease-associated cell content,
- e) comparing said methylation level to that of corresponding reference tissue.
Method for predicting treatment response or prognosis of disease for an individual, characterized by specific methylation levels of one or more methylation variable genomic DNA positions in a disease-associated cell of a tissue sample, comprising:
- a) obtaining a tissue sample comprising genomic DNA having one or more methylation variable positions in one or more regions thereof,
- b) determining the disease-associated cell content within the tissue sample in a quantitative or semi-quantitative manner,
- c) measuring the total DNA methylation level of one or more methylation variable genomic DNA positions of the tissue sample,
- d) determining the DNA methylation level of one or more methylation variable genomic DNA positions within the disease-associated cells from the total DNA methylation level and the disease-associated cell content,
- e) comparing said methylation level to that of corresponding reference tissue.
Kit, comprising
- a) reagents for quantitative or semiquantitative determination of the cell content of a subpopulation of cells within a tissue sample,
- b) reagents for measuring the total DNA methylation level of one or more specific CpG positions of said subpopulation of cells within the tissue sample, and
- c) a container.
Kit, comprising one or more of the following:
- a container,
- one or more primer suitable for the amplification of a subpopulation of cells specific DNA methylation marker to determine the cell content of said subpopulation of cells,
- one or more primer suitable for the amplification of one or more fragments to determine the DNA methylation level of one or more specific CpG positions within the total DNA, and
- operator instructions and/or algorithms to determine the methylation level within the tumor cells of a given sample.
Dependencies are inferred from the claim text.