Background Risk stratification as a routine part of the NHS Breast Screening Programme (NHSBSP) c... more Background Risk stratification as a routine part of the NHS Breast Screening Programme (NHSBSP) could provide a better balance of benefits and harms. We developed BC-Predict, to offer women when invited to the NHSBSP, which collects standard risk factor information; mammographic density; and in a sub-sample, a Polygenic Risk Score (PRS). Methods Risk prediction was estimated primarily from self-reported questionnaires and mammographic density using the Tyrer–Cuzick risk model. Women eligible for NHSBSP were recruited. BC-Predict produced risk feedback letters, inviting women at high risk (≥8% 10-year) or moderate risk (≥5–<8% 10-year) to have appointments to discuss prevention and additional screening. Results Overall uptake of BC-Predict in screening attendees was 16.9% with 2472 consenting to the study; 76.8% of those received risk feedback within the 8-week timeframe. Recruitment was 63.2% with an onsite recruiter and paper questionnaire compared to <10% with BC-Predict onl...
Background The luminal progenitor (LP) population in the normal breast is under the control of pa... more Background The luminal progenitor (LP) population in the normal breast is under the control of paracrine progesterone signalling and likely represents the cell of origin of estrogen receptor negative (ER-) BC. Exogenous progestins, as contraception or menopausal hormone replacement therapy (HRT), increase the risk of ER- and ER+ BC. We sought to examine the effect antagonism of progesterone signalling on the tissue composition and cellular hierarchy of normal human breast and thus the potential for progesterone receptor antagonism in breast cancer prevention. Methods BC-APPS1 is a single arm phase 2 pilot study in which premenopausal women at increased familial BC risk underwent vacuum assisted biopsy (VAB) in the luteal phase of the menstrual cycle prior to commencing a 12 week course of the selective progesterone receptor modulator (SPRM) ulipristal acetate (UA; 5mg daily). VAB was repeated in the final week of therapy. The primary endpoint of BC-APPS1 was change in epithelial Ki67 assessed by immunohistochemistry. Secondary endpoints included toxicity and additional tissue endpoints including immunofluorescence (IF) staining of LP markers, 2D mixed luminal/basal and 3D (mammosphere) colony formation assays, LP fraction by FACS (CD49f+/EPCAM+ cells), single cell and bulk RNAseq, epithelium/stromal laser capture microdissection-based proteomics and tissue stiffness analyses. Baseline samples were compared to a historic cohort of normal risk (n=25) and high risk (n=41) samples for IF analyses. Results Between 03/2016 and 03/2019 26 women were recruited to BC-APPS1 and 24 underwent paired biopsies. The trial met its primary endpoint with a significant reduction in median Ki67 between baseline (4.89%; IQR 4.36-10.42) and 12 weeks (2.41%; IQR 1.57-3.24; p<0.0001). Study procedures were well tolerated with minimal drug toxicity (all G1/2). 2 women discontinued UA due to anxiety: 1 drug induced and 1 induced following development of a small haematoma following VAB1. FACS analysis (n=17) demonstrated significant reduction in the LP proportion with UA treatment (median baseline 44.7% IQR 28.1-55.2 and 12 weeks 25.4% IQR 17.2-37.8; p<0.01). Breast tissue from high-risk women had increased expression of PR+ (10.8% vs 4.5%; p<0.01) and dual Sox9+Ki67+ cells (4.8% vs 0.8%, p<0.01). The Sox9+Ki67+ population reduced significantly with UA therapy in BC-APPS1 (4.4% vs 1.3%, p<0.05). In functional analyses both mammosphere and mixed luminal/basal colony formation reduced with UA treatment and single cell RNAseq (n=8 pairs) and epithelium/stromal laser capture microdissection-based proteomics (n=5 pairs) identified stromal components and remodelling as key pathways perturbed by UA treatment. Tissue stiffness, assessed by atomic force microscopy and previously shown to be positively associated with %mammographic density, was significantly reduced with UA treatment. Conclusions: High risk women have increased surrogate markers of BC risk including proliferating luminal progenitor cells which can be reduced by short term SPRM treatment with UA. Treatment is generally well tolerated and SPRM therapy is an attractive candidate for BC prevention. Longer term studies are warranted and stromal remodelling and breast tissue stiffness data suggest that mammographic density should be investigated as a potential surrogate biomarker of activity. Citation Format: Sacha Howell, Alice Greenhalgh, Robert Pedley, Suad Alghamdi, Amanda Caruso, Mujtaba Ansari, Tiago Moreira, Sue Astlet, Anthony Maxwell, Yit Lim, Hayley Brookes, Faiza Idries, Anthony Howell, D Gareth Evans, Michael Sherratt, Andrew Gilmore, Elaine Harkness, Walid Khaled, Alecia-Jane Twigger, Matt Roberts, Robert Clarke, Bruno Simoes. Results from the breast cancer - anti progestin prevention study 1 (BC-APPS1) trial - a novel approach in breast cancer prevention [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P1-10-01.
The composition of the tumour microenvironment in follicular lymphoma (FL) is a relevant factor i... more The composition of the tumour microenvironment in follicular lymphoma (FL) is a relevant factor in determining disease progression and treatment response. This dataset is a collection of 349 FL diagnostic tissue micro-array (TMA) cores from 130 patients, stained using multi-plex immunofluorescence for: • CD4+ cells • Cytotoxic T cells (CD8+) • T regulatory cells (Tregs [CD4+FOXP3+]) • Macrophages (CD68+) • PD1+ lymphocytes • B cells/follicular dendritic cells (CD21+) • DAPI (4′,6-diamidino-2-phenylindole) nuclear counterstain Changes from previous version 2 ---------- - Raw .im3 image files provided - Spectral library images provided - Labels of nuclear annotations in 16bit format Cohort -------- FL patients according to the WHO 2008 classification were identified from the archives of The Christie NHS Foundation Trust, Manchester, UK. The study was conducted with approval by the North-West Multi-centre Ethics Committee (03/08/016) and according to the Declaration of Helsinki. Examin...
Density residual: description of methods. Appendix 2: Table S1. Univariate and multivariate perfo... more Density residual: description of methods. Appendix 2: Table S1. Univariate and multivariate performance of breast density and the Tyrer-Cuzick and Gail risk models, subgroup analysis by time of cancer diagnosis. (PDF 396 kb)
Background A decrease in breast density due to tamoxifen preventive therapy might indicate greate... more Background A decrease in breast density due to tamoxifen preventive therapy might indicate greater benefit from the drug. It is not known whether mammographic density continues to decline after 1 year of therapy, or whether measures of breast density change are sufficiently stable for personalised recommendations. Methods Mammographic density was measured annually over up to 5 years in premenopausal women with no previous diagnosis of breast cancer but at increased risk of breast cancer attending a family-history clinic in Manchester, UK (baseline 2010-2013). Tamoxifen (20 mg/day) for prevention was prescribed for up to 5 years in one group; the other group did not receive tamoxifen and were matched by age. Fully automatic methods were used on mammograms over the 5-year follow-up: three area-based measures (NN-VAS, Stratus, Densitas) and one volumetric (Volpara). Additionally, percentage breast density at baseline and first follow-up mammograms was measured visually. The size of den...
BACKGROUNDProtein truncating variants in ATM, BRCA1, BRCA2, CHEK2 and PALB2 are associated with i... more BACKGROUNDProtein truncating variants in ATM, BRCA1, BRCA2, CHEK2 and PALB2 are associated with increased breast cancer risk, but risks associated with missense variants in these genes are uncertain.METHODSCombining 59,639 breast cancer cases and 53,165 controls, we sampled training (80%) and validation (20%) sets to analyze rare missense variants in ATM (1,146 training variants), BRCA1 (644), BRCA2 (1,425), CHEK2 (325) and PALB2 (472). We evaluated breast cancer risks according to five in-silico prediction-of-deleteriousness algorithms, functional protein domain, and frequency, using logistic regression models and also mixture models in which a subset of variants was assumed to be risk-associated.RESULTSThe most predictive in-silico algorithms were Helix (BRCA1, BRCA2 and CHEK2) and CADD (ATM). Increased risks appeared restricted to functional protein domains for ATM (FAT and PIK domains) and BRCA1 (RING and BRCT domains). For ATM, BRCA1 and BRCA2, data were compatible with small s...
Background: Genome-wide association studies have identified over 170 common breast cancer suscept... more Background: Genome-wide association studies have identified over 170 common breast cancer susceptibility loci, many of them with differential associations by estrogen receptor (ER). How these variants are related to other tumor features is unclear. Methods: Analyses included 106,571 invasive breast cancer cases and 95,762 controls of European ancestry with data on 178 genotyped or imputed single nucleotide polymorphisms (SNPs). We used two-stage polytomous logistic regression models to evaluate SNPs in relation to multiple tumor features (ER, progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and grade) adjusting for each other, and to intrinsic-like subtypes. Results: Nearly half of the SNPs (85 of 178) were associated with at least one tumor feature (false discovery rate <5%). Case-case comparisons identified ER and grade as the most common heterogeneity sources, followed by PR and HER2. Case-control comparisons among these 85 SNPs with intrinsic-like s...
While the requirement for thresholds for testing for mutations in BRCA1/2 is being questioned, th... more While the requirement for thresholds for testing for mutations in BRCA1/2 is being questioned, they are likely to remain for individuals unaffected by a relevant cancer. It is still useful to provide pretesting likelihoods, but models need to take into account tumour pathology. The Manchester Scoring System (MSS) is a well-used, simple, paper-based model for assessing carrier probability that already incorporates pathology data. We have used mutation testing data from 4115 unrelated samples from affected non-Jewish individuals alongside tumour pathology to further refine the scoring system. Adding additional points for high-grade serous ovarian cancer <60 (HGSOC=+2) and adding grade score to those with triple-negative breast cancer, while reducing the score for those with HER2+ breast cancer (-6), resulted in significantly improved sensitivity and minor improvements in specificity to the MSS. Sporadic HGSOC <60 years thus reached a score of 15-19 points within the 10% grouping...
This study compared mammographic density over time between women who developed breast cancer case... more This study compared mammographic density over time between women who developed breast cancer cases and women who did not controls. Cases had an initial negative mammographic screen and another three years later when cancer was diagnosed. Cases were matched to three controls with two successive negative screens by age, year of mammogram, BMI, parity, menopausal status and HRT use. Mammographic density was measured by VolparaTM. There was a significant reduction in percentage density in the affected breast for cases 5.2 to 4.8i?ź%, pi?ź<i?ź0.001 and for the same matched breast in controls 4.9 to 4.5, pi?ź<i?ź0.001. Similar results were found for the unaffected breast. After adjusting for density measures at the initial screen, case-control status was only significantly associated with fibroglandular volume in the unaffected breast adjusted mean 45.8i?źcm3 in cases, 44.0i?źcm3 in controls, pi?ź=i?ź0.008. The results suggest changes in mammographic density may be less important than initial mammographic density.
This study investigates variations in mammographic density by ethnic group in women attending the... more This study investigates variations in mammographic density by ethnic group in women attending the NHS breast screening programme in Greater Manchester. Density was estimated using VolparaTM and QuantraTM. Data was analysed for 651 Asian/Asian British, 416 Black/Black British, 394 Jewish origin, 181 'Mixed', 700 'Other' and a random sample of 10,000 women who declared their ethnic origin as White British or Irish. Age ranged from 46---84 years and mean BMI was 27.4i¾?kg/m2. Fibroglandular volume VolparaTM was highest in women of Black/Black British origin 59.4i¾?cm3 and lowest in Asian/Asian British women 47.9i¾?cm3. After adjusting for a number of hormonal and other factors the magnitude of the difference between groups decreased, however, there were still a number of statistical differences between groups. Ethnic differences in mammographic density and personal factors may subsequently contribute to differences in breast cancer incidence.
Background Risk stratification as a routine part of the NHS Breast Screening Programme (NHSBSP) c... more Background Risk stratification as a routine part of the NHS Breast Screening Programme (NHSBSP) could provide a better balance of benefits and harms. We developed BC-Predict, to offer women when invited to the NHSBSP, which collects standard risk factor information; mammographic density; and in a sub-sample, a Polygenic Risk Score (PRS). Methods Risk prediction was estimated primarily from self-reported questionnaires and mammographic density using the Tyrer–Cuzick risk model. Women eligible for NHSBSP were recruited. BC-Predict produced risk feedback letters, inviting women at high risk (≥8% 10-year) or moderate risk (≥5–<8% 10-year) to have appointments to discuss prevention and additional screening. Results Overall uptake of BC-Predict in screening attendees was 16.9% with 2472 consenting to the study; 76.8% of those received risk feedback within the 8-week timeframe. Recruitment was 63.2% with an onsite recruiter and paper questionnaire compared to <10% with BC-Predict onl...
Background The luminal progenitor (LP) population in the normal breast is under the control of pa... more Background The luminal progenitor (LP) population in the normal breast is under the control of paracrine progesterone signalling and likely represents the cell of origin of estrogen receptor negative (ER-) BC. Exogenous progestins, as contraception or menopausal hormone replacement therapy (HRT), increase the risk of ER- and ER+ BC. We sought to examine the effect antagonism of progesterone signalling on the tissue composition and cellular hierarchy of normal human breast and thus the potential for progesterone receptor antagonism in breast cancer prevention. Methods BC-APPS1 is a single arm phase 2 pilot study in which premenopausal women at increased familial BC risk underwent vacuum assisted biopsy (VAB) in the luteal phase of the menstrual cycle prior to commencing a 12 week course of the selective progesterone receptor modulator (SPRM) ulipristal acetate (UA; 5mg daily). VAB was repeated in the final week of therapy. The primary endpoint of BC-APPS1 was change in epithelial Ki67 assessed by immunohistochemistry. Secondary endpoints included toxicity and additional tissue endpoints including immunofluorescence (IF) staining of LP markers, 2D mixed luminal/basal and 3D (mammosphere) colony formation assays, LP fraction by FACS (CD49f+/EPCAM+ cells), single cell and bulk RNAseq, epithelium/stromal laser capture microdissection-based proteomics and tissue stiffness analyses. Baseline samples were compared to a historic cohort of normal risk (n=25) and high risk (n=41) samples for IF analyses. Results Between 03/2016 and 03/2019 26 women were recruited to BC-APPS1 and 24 underwent paired biopsies. The trial met its primary endpoint with a significant reduction in median Ki67 between baseline (4.89%; IQR 4.36-10.42) and 12 weeks (2.41%; IQR 1.57-3.24; p<0.0001). Study procedures were well tolerated with minimal drug toxicity (all G1/2). 2 women discontinued UA due to anxiety: 1 drug induced and 1 induced following development of a small haematoma following VAB1. FACS analysis (n=17) demonstrated significant reduction in the LP proportion with UA treatment (median baseline 44.7% IQR 28.1-55.2 and 12 weeks 25.4% IQR 17.2-37.8; p<0.01). Breast tissue from high-risk women had increased expression of PR+ (10.8% vs 4.5%; p<0.01) and dual Sox9+Ki67+ cells (4.8% vs 0.8%, p<0.01). The Sox9+Ki67+ population reduced significantly with UA therapy in BC-APPS1 (4.4% vs 1.3%, p<0.05). In functional analyses both mammosphere and mixed luminal/basal colony formation reduced with UA treatment and single cell RNAseq (n=8 pairs) and epithelium/stromal laser capture microdissection-based proteomics (n=5 pairs) identified stromal components and remodelling as key pathways perturbed by UA treatment. Tissue stiffness, assessed by atomic force microscopy and previously shown to be positively associated with %mammographic density, was significantly reduced with UA treatment. Conclusions: High risk women have increased surrogate markers of BC risk including proliferating luminal progenitor cells which can be reduced by short term SPRM treatment with UA. Treatment is generally well tolerated and SPRM therapy is an attractive candidate for BC prevention. Longer term studies are warranted and stromal remodelling and breast tissue stiffness data suggest that mammographic density should be investigated as a potential surrogate biomarker of activity. Citation Format: Sacha Howell, Alice Greenhalgh, Robert Pedley, Suad Alghamdi, Amanda Caruso, Mujtaba Ansari, Tiago Moreira, Sue Astlet, Anthony Maxwell, Yit Lim, Hayley Brookes, Faiza Idries, Anthony Howell, D Gareth Evans, Michael Sherratt, Andrew Gilmore, Elaine Harkness, Walid Khaled, Alecia-Jane Twigger, Matt Roberts, Robert Clarke, Bruno Simoes. Results from the breast cancer - anti progestin prevention study 1 (BC-APPS1) trial - a novel approach in breast cancer prevention [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P1-10-01.
The composition of the tumour microenvironment in follicular lymphoma (FL) is a relevant factor i... more The composition of the tumour microenvironment in follicular lymphoma (FL) is a relevant factor in determining disease progression and treatment response. This dataset is a collection of 349 FL diagnostic tissue micro-array (TMA) cores from 130 patients, stained using multi-plex immunofluorescence for: • CD4+ cells • Cytotoxic T cells (CD8+) • T regulatory cells (Tregs [CD4+FOXP3+]) • Macrophages (CD68+) • PD1+ lymphocytes • B cells/follicular dendritic cells (CD21+) • DAPI (4′,6-diamidino-2-phenylindole) nuclear counterstain Changes from previous version 2 ---------- - Raw .im3 image files provided - Spectral library images provided - Labels of nuclear annotations in 16bit format Cohort -------- FL patients according to the WHO 2008 classification were identified from the archives of The Christie NHS Foundation Trust, Manchester, UK. The study was conducted with approval by the North-West Multi-centre Ethics Committee (03/08/016) and according to the Declaration of Helsinki. Examin...
Density residual: description of methods. Appendix 2: Table S1. Univariate and multivariate perfo... more Density residual: description of methods. Appendix 2: Table S1. Univariate and multivariate performance of breast density and the Tyrer-Cuzick and Gail risk models, subgroup analysis by time of cancer diagnosis. (PDF 396 kb)
Background A decrease in breast density due to tamoxifen preventive therapy might indicate greate... more Background A decrease in breast density due to tamoxifen preventive therapy might indicate greater benefit from the drug. It is not known whether mammographic density continues to decline after 1 year of therapy, or whether measures of breast density change are sufficiently stable for personalised recommendations. Methods Mammographic density was measured annually over up to 5 years in premenopausal women with no previous diagnosis of breast cancer but at increased risk of breast cancer attending a family-history clinic in Manchester, UK (baseline 2010-2013). Tamoxifen (20 mg/day) for prevention was prescribed for up to 5 years in one group; the other group did not receive tamoxifen and were matched by age. Fully automatic methods were used on mammograms over the 5-year follow-up: three area-based measures (NN-VAS, Stratus, Densitas) and one volumetric (Volpara). Additionally, percentage breast density at baseline and first follow-up mammograms was measured visually. The size of den...
BACKGROUNDProtein truncating variants in ATM, BRCA1, BRCA2, CHEK2 and PALB2 are associated with i... more BACKGROUNDProtein truncating variants in ATM, BRCA1, BRCA2, CHEK2 and PALB2 are associated with increased breast cancer risk, but risks associated with missense variants in these genes are uncertain.METHODSCombining 59,639 breast cancer cases and 53,165 controls, we sampled training (80%) and validation (20%) sets to analyze rare missense variants in ATM (1,146 training variants), BRCA1 (644), BRCA2 (1,425), CHEK2 (325) and PALB2 (472). We evaluated breast cancer risks according to five in-silico prediction-of-deleteriousness algorithms, functional protein domain, and frequency, using logistic regression models and also mixture models in which a subset of variants was assumed to be risk-associated.RESULTSThe most predictive in-silico algorithms were Helix (BRCA1, BRCA2 and CHEK2) and CADD (ATM). Increased risks appeared restricted to functional protein domains for ATM (FAT and PIK domains) and BRCA1 (RING and BRCT domains). For ATM, BRCA1 and BRCA2, data were compatible with small s...
Background: Genome-wide association studies have identified over 170 common breast cancer suscept... more Background: Genome-wide association studies have identified over 170 common breast cancer susceptibility loci, many of them with differential associations by estrogen receptor (ER). How these variants are related to other tumor features is unclear. Methods: Analyses included 106,571 invasive breast cancer cases and 95,762 controls of European ancestry with data on 178 genotyped or imputed single nucleotide polymorphisms (SNPs). We used two-stage polytomous logistic regression models to evaluate SNPs in relation to multiple tumor features (ER, progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and grade) adjusting for each other, and to intrinsic-like subtypes. Results: Nearly half of the SNPs (85 of 178) were associated with at least one tumor feature (false discovery rate <5%). Case-case comparisons identified ER and grade as the most common heterogeneity sources, followed by PR and HER2. Case-control comparisons among these 85 SNPs with intrinsic-like s...
While the requirement for thresholds for testing for mutations in BRCA1/2 is being questioned, th... more While the requirement for thresholds for testing for mutations in BRCA1/2 is being questioned, they are likely to remain for individuals unaffected by a relevant cancer. It is still useful to provide pretesting likelihoods, but models need to take into account tumour pathology. The Manchester Scoring System (MSS) is a well-used, simple, paper-based model for assessing carrier probability that already incorporates pathology data. We have used mutation testing data from 4115 unrelated samples from affected non-Jewish individuals alongside tumour pathology to further refine the scoring system. Adding additional points for high-grade serous ovarian cancer <60 (HGSOC=+2) and adding grade score to those with triple-negative breast cancer, while reducing the score for those with HER2+ breast cancer (-6), resulted in significantly improved sensitivity and minor improvements in specificity to the MSS. Sporadic HGSOC <60 years thus reached a score of 15-19 points within the 10% grouping...
This study compared mammographic density over time between women who developed breast cancer case... more This study compared mammographic density over time between women who developed breast cancer cases and women who did not controls. Cases had an initial negative mammographic screen and another three years later when cancer was diagnosed. Cases were matched to three controls with two successive negative screens by age, year of mammogram, BMI, parity, menopausal status and HRT use. Mammographic density was measured by VolparaTM. There was a significant reduction in percentage density in the affected breast for cases 5.2 to 4.8i?ź%, pi?ź<i?ź0.001 and for the same matched breast in controls 4.9 to 4.5, pi?ź<i?ź0.001. Similar results were found for the unaffected breast. After adjusting for density measures at the initial screen, case-control status was only significantly associated with fibroglandular volume in the unaffected breast adjusted mean 45.8i?źcm3 in cases, 44.0i?źcm3 in controls, pi?ź=i?ź0.008. The results suggest changes in mammographic density may be less important than initial mammographic density.
This study investigates variations in mammographic density by ethnic group in women attending the... more This study investigates variations in mammographic density by ethnic group in women attending the NHS breast screening programme in Greater Manchester. Density was estimated using VolparaTM and QuantraTM. Data was analysed for 651 Asian/Asian British, 416 Black/Black British, 394 Jewish origin, 181 'Mixed', 700 'Other' and a random sample of 10,000 women who declared their ethnic origin as White British or Irish. Age ranged from 46---84 years and mean BMI was 27.4i¾?kg/m2. Fibroglandular volume VolparaTM was highest in women of Black/Black British origin 59.4i¾?cm3 and lowest in Asian/Asian British women 47.9i¾?cm3. After adjusting for a number of hormonal and other factors the magnitude of the difference between groups decreased, however, there were still a number of statistical differences between groups. Ethnic differences in mammographic density and personal factors may subsequently contribute to differences in breast cancer incidence.
Uploads
Papers by Elaine Harkness