BRCA1 Gene MutationNCT01907438
Transformation Potential of E2 Exposed Breast Cancer Susceptibility Gene Mutation Heterozygous Epithelial Breast Cells
An observational study of BRCA1 Gene Mutation and BRCA2 Gene Mutation, run by Hadassah Medical Organization at 1 site in Israel.
Trial 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. Read more about this study
- Who is it for?
- Ages 25 to 50 years, women onlyHealthy volunteers can take part
- Conditions the study is for
- BRCA1 Gene Mutation, BRCA2 Gene Mutation
- How many people
- 30 participants
- Where
- 1 siteIsrael
- Study dates
- Sep 2013to Sep 2015
- How long will I take part?
- 7 Days
- Phase
- Observational
- Are there results?
- None posted in the registry
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 25 to 50 years
- Women only
- Healthy volunteers accepted
- Adult
Study population: human breast tissue samples will be obtained from risk-reducing mastectomy in healthy premenopausal women with BRCA1 and BRCA2 mutations. Tissues from premenopausal women undergoing esthetic breast surgery with no family history of breast or ovarian cancers also will be obtained.
Who may be able to join
- healthy
- premenopausal
- women
- BRCA1 mutations
- BRCA2 mutations
Who cannot join
- women with breast or ovarian cancer
Trial locations and contacts
1 site in 1 country.
Asher Y Salmon, MD, PhD contact
Israel1 site
Hadassah medical center
Jerusalem
Tamar Zahavi, PhD, tamar.harel1@mail.huji.ac.il
Asher Y Salmon, MD, PhD
About this study
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
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.
Read the 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 Hank's + 2% fetal bovine serum (FBS) and 0.1 mg/mL 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.
- Conditions
- BRCA1 Gene Mutation, BRCA2 Gene Mutation
- Keywords
- BRCA, estrogen, epithelial breast cells, genome profile
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
| Study group | Role | What they receive |
|---|---|---|
| non-carriers | Tissues from premenopausal women undergoing esthetic breast surgery with no family history of breast or ovarian cancers will be obtained.epithelial breast cells will be isolated, treated with estrogen for 48 h and then will be irradiated. | |
| BRCA1 mutation carriers | Tissues from risk-reducing mastectomy in healthy premenopausal women with BRCA1 mutations undergoing.epithelial breast cells will be isolated, treated with estrogen for 48 h and then will be irradiated. | |
| BRCA2 mutation carriers | Tissues from risk-reducing mastectomy in healthy premenopausal women with BRCA2 mutations undergoing.epithelial breast cells will be isolated, treated with estrogen for 48 h and then will be irradiated. |
What is being measured
The primary outcome is the main question the study is designed to answer.
The transcriptional profile of heterozygous epithelial breast cells
we will perform a RNA-seq expression profiling analysis of normal estrogen exposed BRCA heterozygous epithelial cells following irradiation.
Measured: human breast tissue samples will be obtained from premenopausal women undergoing breast surgery. cells will be isolated immediatly and 48h later the transcriptional profile will be identified.
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted25 Jul 2013 (estimated)
- Study startSep 2013
- Primary completionSep 2014 (estimated)
- Study completionSep 2015 (estimated)
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Other trials for BRCA1 Gene MutationStudies of the same condition, open and under way.
- More trials run by Hadassah Medical OrganizationThe sponsor's other registered studies.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Observational
- Observational model
- Case-control
- Time perspective
- Cross-sectional
- Enrolment
- 30 participants (estimated)
- Target duration
- 7 Days
Sponsor and investigators
- Lead sponsor
- Hadassah Medical Organization
- Sponsor type
- University, hospital or other
- Responsible party
- Principal investigator, Asher Salmon, Hadassah Medical Organization
- Investigators
- Asher Y Salmon, MD, PhD, Principal investigator, Hadassah Medical Organization
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT01907438
- Sponsor's protocol ID
- ISF-1702/12
- Status last verified
- Jul 2013
- First submitted
- 22 Jul 2013
- Last updated
- 25 Jul 2013
- Results posted
- No
- Data monitoring committee
- Yes
Study at a glance
From the registry record, last updated by the sponsor on 25 Jul 2013. Recruitment status is as the sponsor reported it: confirm with the study team.
- 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
- ClinicalTrials.gov ID
- NCT01907438
- Acronym / sponsor ID
- ISF-1702/12
- Conditions
- BRCA1 Gene Mutation, BRCA2 Gene Mutation
- Keywords
- BRCA, estrogen, epithelial breast cells, genome profile
- Study type
- Observational
- Recruitment status
- Unknown status
- Enrolment
- 30 participants (estimated)
- Lead sponsor
- Hadassah Medical OrganizationUniversity, hospital or other
- Study start
- Sep 2013
- Primary completion
- Sep 2014 (estimated)
- Study completion
- Sep 2015 (estimated)
- First posted
- 25 Jul 2013 (estimated)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
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.
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 Hank's + 2% fetal bovine serum (FBS) and 0.1 mg/mL 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.
Study design
- Observational model
- Case-control
- Time perspective
- Cross-sectional
- Target follow-up
- 7 Days
- Number of arms
- 3
| Arm | Type | Receives |
|---|---|---|
| non-carriers | – | –Tissues from premenopausal women undergoing esthetic breast surgery with no family history of breast or ovarian cancers will be obtained.epithelial breast cells will be isolated, treated with estrogen for 48 h and then will be irradiated. |
| BRCA1 mutation carriers | – | –Tissues from risk-reducing mastectomy in healthy premenopausal women with BRCA1 mutations undergoing.epithelial breast cells will be isolated, treated with estrogen for 48 h and then will be irradiated. |
| BRCA2 mutation carriers | – | –Tissues from risk-reducing mastectomy in healthy premenopausal women with BRCA2 mutations undergoing.epithelial breast cells will be isolated, treated with estrogen for 48 h and then will be irradiated. |
Who may qualify
- Ages
- 25 to 50 years
- Sex
- Female only
- Healthy volunteers
- Accepted
- Age groups
- Adult
- Study population
- human breast tissue samples will be obtained from risk-reducing mastectomy in healthy premenopausal women with BRCA1 and BRCA2 mutations. Tissues from premenopausal women undergoing esthetic breast surgery with no family history of breast or ovarian cancers also will be obtained.
Key inclusion criteria (5 listed)
- healthy
- premenopausal
- women
- BRCA1 mutations
- BRCA2 mutations
Key exclusion criteria (1 listed)
- women with breast or ovarian cancer
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Primary outcome
The transcriptional profile of heterozygous epithelial breast cells
Time frame: human breast tissue samples will be obtained from premenopausal women undergoing breast surgery. cells will be isolated immediatly and 48h later the transcriptional profile will be identified.
we will perform a RNA-seq expression profiling analysis of normal estrogen exposed BRCA heterozygous epithelial cells following irradiation.
Results and safety
Results are not available in the source record.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 25 Jul 2013.
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Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part. This study is not recruiting at the moment.
Asher Y Salmon, MD, PhD contact
Asher Y Salmon, MD, PhD principal investigator
Hadassah Medical Organization
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
An observational study of BRCA1 Gene Mutation and BRCA2 Gene Mutation, run by Hadassah Medical Organization at 1 site in Israel.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 5 inclusion and 1 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: status not confirmed, as of the sponsor's last update on 25 Jul 2013. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in Israel. Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
Results are not available in the source record.
How can a doctor refer a patient or contact the research team?
Contact Asher Y Salmon, MD, PhD (asalmon@hadassah.org.il), or a site near the patient. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
The transcriptional profile of heterozygous epithelial breast cells (time frame: human breast tissue samples will be obtained from premenopausal women undergoing breast surgery. cells will be isolated immediatly and 48h later the transcriptional profile will be identified.). It is what the study was designed to measure, not a finding.
Where can I read the original study information?
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 25 Jul 2013.