Ignite Creation Date:
2024-05-06 @ 1:48 AM
Last Modification Date:
2024-10-26 @ 11:10 AM
Study NCT ID:
NCT01907438
Status:
UNKNOWN
Last Update Posted:
2013-07-25
First Post:
2013-07-22
Brief Title:
Transformation Potential of E2 Exposed Breast Cancer Susceptibility Gene Mutation Heterozygous Epithelial Breast Cells
Sponsor:
Hadassah Medical Organization
Organization:
Hadassah Medical Organization
Study Overview
Official Title:
Identification of the Transformation Potential of Normal Estrogen Exposed BRCA1 Breast Cancer Susceptibility Gene 1 and BRCA2 Breast Cancer Susceptibility Gene 2 Heterozygous Epithelial Breast Cells Due to Irradiation
Status:
UNKNOWN
Status Verified Date:
2013-07
Last Known Status:
NOT_YET_RECRUITING
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
Susceptibility to breast cancer is related to the combination of genetic hormonal and multiple other environmental risk factors such as mutations in the BRCA gene and excess exposure to exogenous estrogen respectively BRCA is a nuclear protein that maintains genome stability by acting as a key player in the DNA repair complex Recently evidence has emerged that BRCA mutation heterozygosis itself enhances aborted DNA repair and can contribute to breast cancer initiation after exposure to irradiation In our preliminary results on short-term lymphocyte cultures we found additional evidence that healthy heterozygous BRCA1 and BRCA2 mutation carriers have a different response to DNA damage than do non-carriers
The main aim of our ongoing project is to identify the transcriptional modulation and transformation potential of normal BRCA1 and BRCA2 mutation heterozygous epithelial breast cells following irradiation and to examine how it is affected by exposure to estrogen Our hypotheses will be investigated by RNA-seq and microRNA-seq in order to identify a unique molecular expression profile of the estrogen exposed cells following ionizing irradiation
Understanding the role of BRCA heterozygosity in cell response to exposure to estrogen and to irradiation may facilitate the development of more appropriate diagnostic and therapeutic strategies for these individuals
The main aim of our ongoing project is to identify the transcriptional modulation and transformation potential of normal BRCA1 and BRCA2 mutation heterozygous epithelial breast cells following irradiation and to examine how it is affected by exposure to estrogen Our hypotheses will be investigated by RNA-seq and microRNA-seq in order to identify a unique molecular expression profile of the estrogen exposed cells following ionizing irradiation
Understanding the role of BRCA heterozygosity in cell response to exposure to estrogen and to irradiation may facilitate the development of more appropriate diagnostic and therapeutic strategies for these individuals
Detailed Description:
To study the different cell lineages we plan to isolate and propagate luminal progenitors from a normal human breast tissue To this end 10 human breast tissue samples will be obtained from risk-reducing mastectomy in healthy premenopausal women with BRCA1 mutations Tissues from 10 premenopausal women undergoing esthetic breast surgery with no family history of breast or ovarian cancers will serve as a control group In order to isolate primary epithelial cells human mammary tissue will be minced and enzymatically digested overnight in collagenase and hyaluronidase to yield suspension of epithelial organoids These organoids will be collected and further digested with trypsin dispase and deoxyribonuclease 1 DNAse will be filtered to generate a single cell suspension resuspended in Hanks 2 fetal bovine serum FBS and 01 mgmL DNAse and also incubated with a blocking antibody for 15 minutes on ice
Cells will be treated with estrogen E2 10 nM for 48 h and then will be irradiated for inducing double-strand break Cells will be irradiated with 8 Gray Gy using a Co60 source
To accomplish our study estrogen exposed and unexposed BRCA mutation heterozygous epithelial breast cells will be irradiated and 1 h later RNA will be extracted from the cells using Tri-reagent Sigma The RNA will be converted to a library of cDNA complementary DNA fragments and will be sent for deep sequencing in the illumina Hi-Seq platform
Cells will be treated with estrogen E2 10 nM for 48 h and then will be irradiated for inducing double-strand break Cells will be irradiated with 8 Gray Gy using a Co60 source
To accomplish our study estrogen exposed and unexposed BRCA mutation heterozygous epithelial breast cells will be irradiated and 1 h later RNA will be extracted from the cells using Tri-reagent Sigma The RNA will be converted to a library of cDNA complementary DNA fragments and will be sent for deep sequencing in the illumina Hi-Seq platform
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
None
Is a FDA Regulated Device?:
None
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None