Colonic polyps may arise from BRAF inhibitor treatment of melanoma, possibly due to paradoxical activation of the mitogen-activated protein (MAP)-kinase pathway. In an alternative evidence based scenario, tubular colo...Colonic polyps may arise from BRAF inhibitor treatment of melanoma, possibly due to paradoxical activation of the mitogen-activated protein (MAP)-kinase pathway. In an alternative evidence based scenario, tubular colonic adenomas with APC gene mutations have also been identified in the context of BRAF inhibitor treatment, in the absence of mutations of MAPK genes. A minority of colorectal cancers develop by an alternative “serrated polyp pathway”. This article postulates a novel hypothesis, that the established phenotypic and molecular characteristics of serrated colonic polyps/CRC offer an intriguing insight into the pathobiology of BRAF inhibitor induced colonic polyps. Serrated polyps are characterized by a CpG island methylation phenotype, MLH1 silencing and cellular senescence. They also have BRAF mutations. The contention is that BRAF inhibitor induced polyps mimic the afore-described histology and molecular features of serrated polyps with the exception that instead of the presence of BRAF mutations they induce C-RAF homodimers and B-RAF: C-RAF heterodimers.展开更多
Ameloblastoma is a benign but locally aggressive odontogenic neoplasm that accounts for 10% of all tumors arising in the mandible and maxilla (1). Eighty percent of ameloblastomas arise in the mandible, and they are u...Ameloblastoma is a benign but locally aggressive odontogenic neoplasm that accounts for 10% of all tumors arising in the mandible and maxilla (1). Eighty percent of ameloblastomas arise in the mandible, and they are usually found in young adults. It frequently recurs if not adequately展开更多
BRAF is a serine/threonine kinase that harbors activating mutations in^7%of human malignancies and^60%of melanomas.Despite initial clinical responses to BRAF inhibitors,patients frequently develop drug resistance.To i...BRAF is a serine/threonine kinase that harbors activating mutations in^7%of human malignancies and^60%of melanomas.Despite initial clinical responses to BRAF inhibitors,patients frequently develop drug resistance.To identify candidate therapeutic targets for BRAF inhibitor resistant melanoma,we conduct CRISPR screens in melanoma cells harboring an activating BRAF mutation that had also acquired resistance to BRAF inhibitors.To investigate the mechanisms and pathways enabling resistance to BRAF inhibitors in melanomas,we integrate expression,ATAC-seq,and CRISPR screen data.We identify the JUN family transcription factors and the ETS family transcription factor ETV5 as key regulators of CDK6,which together enable resistance to BRAF inhibitors in melanoma cells.Our findings reveal genes contributing to resistance to a selective BRAF inhibitor PLX4720,providing new insights into gene regulation in BRAF inhibitor resistant melanoma cells.展开更多
Objective:SOX4,a transcription factor,has been found to contribute to tumorigenesis in several cancers.This study was performed to determine whether SOX4 mediates BRAF inhibitor resistance in melanoma.Methods:Melanoma...Objective:SOX4,a transcription factor,has been found to contribute to tumorigenesis in several cancers.This study was performed to determine whether SOX4 mediates BRAF inhibitor resistance in melanoma.Methods:Melanoma cell lines with acquired resistance to BRAF inhibitor(SK-MEL-5R,SK-MEL-28R,and A375R)were generated by adding escalating concentrations of PLX4032 into parental SK-MEL-5,SK-MEL-28,and A375 cells for>6 months.The expression of SOX4 and insulin-like growth factor 1 receptor(IGF-1R)was measured by quantitative real-time polymerase chain reaction(qRT-PCR)and Westem blotting.The downstream signaling of IGF-1R was detected by Westem blotting.SOX4 and IGF-1R overexpression or knockdown was conducted by lentivirus transfection.Cell viability and apoptosis were demonstrated by MTT and flow cytometry,respectively.The binding ability of SOX4 to IGF-1R promoter was determined by chromatin immunoprecipitation quantitative PCR assay.Results:SOX4 was upregulated in BRAF inhibitor-resistant melanoma cells as compared with parental cells(SK-MEL-5 group,1.02 vs.6.33;SK-MEL-28 group,1.03 vs.3.22;A375 group,1.00 vs.1.86;t=°7.069,29.26,and 5.291,respectively;all P<0.01),and PLX4032 treatment could not alter the expression of SOX4 in resistant cells.SOX4 overexpreesion attenuated the response of parental cells to PLX4032(for cell viability,SK-MEL-5 group:77.76%vs.104.28%,F=91.50;SK-MEL-28 group:60.59%vs.93.13%,F=171.8;A375 group:62.50%vs.80.87%,F=47.15.For apoptosis rates,SK-MEL-5 group:34.90%vs.14.31%,F=4.781;SK-MEL-28 group,40.8%vs.29.4%,F=13.32,P=0.063;A375 group:40.20%vs.17.09%,F=11.39;all P<0.05,otherwise indicated).While SOX4 knockdown enhanced the response of resistant cells to PLX4032(for cell viability,SK-MEL-5R group:93.75%vs.69.53%,F=94.45,SK-MEL-28R group:95.60%vs.66.79%,F=30.41,A375R group:95.51%vs.59.98%,F=111.6;for apoptosis rates,SK-MEL-5R group:16.2%vs.44.4%,F=25.67,SK-MEL-28R group:26.59%vs.44.20%,F=158.0,A375R group:5.98%vs.31.51%,F=14.35,and all P<0.01).Chromatin immunoprecipitation quantitative PCR assay demonstrated that SOX4 binded to the promoter of IGF-1R(1.04 vs.1.94[-1044 to-920 bp]and 0.110 vs.0.139[GAPDH],F=534.5,P<0.01).In addition,SOX4 overexpression increased IGF-1R end its downstream phosphorylated ERK,phosphorylated AKT,and phosphorylated STAT3 expression,while SOX4 knockdown exerted the opposite effects.Moreover,IGF-1R knockdown overcame SOX4 overexpreesion-induced PLX4032 resistance(cell viability:35.85%vs.52.79%vs.37.84%[A375 group,negative control group vs.SOX4 overexpressing group vs.SOX4 overexpressing+sh-IGF-1R group];apoptosis rates:25.30%vs.9.56%vs.22.26[A375 group,negative control group vs.SOX4 overexprassing group vs.SOX4 overexpressing+sh-IGF-1R group];F=13.01 and 41.87,respectively;all P<0.01),while IGF-1R overexpression abrogated SOX4 knockdown-induced response enhancement to PLX4032 for comparison of negative control group,sh-SOX4group and sh-SOX4+IGF-1 R overexpreesing group(cell viability:96.62%vs.86.86%vs.97.26%(A375R),98.15%vs.81.63%vs.98.49%[SK-MEL-5R];apoptosis rates:13.81%vs.32.00%vs.12.16[A375R],29.70%vs.41.40%vs.26.10%[SK-MEL-5R];F=13.56,12.86,38.81,and Conclusion:SOX4 mediates BRAF inhibitor resistance in melanoma through regulation of IGF-1R signaling.SOX4 might serve as a potential target for the treatment of BRAF inhibitor-resistant melanoma.39.85,respectively;all P<0.01).展开更多
BACKGROUND Immune checkpoint inhibitors have revolutionized the treatment of patients with unresectable metastatic malignant melanoma.In addition to systemic side effects,several usually mild ocular adverse effects ha...BACKGROUND Immune checkpoint inhibitors have revolutionized the treatment of patients with unresectable metastatic malignant melanoma.In addition to systemic side effects,several usually mild ocular adverse effects have been reported.We report a case of rarely reported vision-threatening bilateral panuveitis with serous retinal detachment,thickened choroid,and chorioretinal folds associated with dabrafenib and trametinib targeted therapy for B-Raf proto-oncogene serine/threonine kinase(BRAF)mutant metastatic cutaneous melanoma.CASE SUMMARY A 59-year-old female patient with metastatic melanoma treated with dabrafenib and trametinib presented with blurry vision and central scotoma lasting for 3 d in both eyes.Clinical examination and multimodal imaging revealed inflammatory cells in the anterior chamber,mild vitritis,bullous multiple serous retinal detachments,and chorioretinal folds in both eyes.Treatment with dabrafenib and trametinib was suspended,and the patient was treated with topical and intravenous corticosteroids followed by oral corticosteroid treatment with a tapering schedule.One and a half months after the disease onset,ocular morphological and functional improvement was noted.Due to the metastatic melanoma dissemination,BRAF/mitogen-activated protein kinase inhibitors were reintroduced and some mild ocular adverse effects reappeared,which later subsided after receiving oral corticosteroids.CONCLUSION Patients on combination therapy with dabrafenib and trametinib may rarely develop severe bilateral panuveitis with a good prognosis.Further studies have to establish potential usefulness of ophthalmological examination for asymptomatic patients.Furthermore,appropriate guidelines for managing panuveitis associated with dabrafenib and trametinib should be established.展开更多
BRAF has been recognized as a promising target for cancer therapy. A number of crystal structures have been published. Molecular docking is one of the most effective techniques in the field of computer-aided drug des...BRAF has been recognized as a promising target for cancer therapy. A number of crystal structures have been published. Molecular docking is one of the most effective techniques in the field of computer-aided drug design（CADD）. Appropriate protein conformation and docking method are essential for the successful virtual screening experiments. One approach considering protein flexibility and multiple docking methods was proposed in this study. Six DFG-in/αC-helix-out crystal structures of BRAF, three docking programs（Glide, GOLD and Ligand Fit） and 12 scoring functions were applied for the best combination by judging from the results of pose prediction and retrospective virtual screening（VS）. The most accurate results（mean RMSD of about 0.6 A） of pose prediction were obtained with two complex structures（PDB： 3 C4 C and 3 SKC） using Glide SP. From the retrospective VS, the most active compounds were identified by using the complex structure of 3 SKC, indicated by a ROC/AUC score of 0.998 and an EF of 20.6 at 5% of the database screen with Glide-SP. On the whole, PDB 3 SKC could achieve a higher rate of correct reproduction, a better enrichment and more diverse compounds. A comparison of 3 SKC and the other X-ray crystal structures led to a rationale for the docking results. PDB 3 SKC could achieve a broad range of sulfonamide substitutions through an expanded hydrophobic pocket formed by a further shift of the αC-helix. Our study emphasized the necessity and significance of protein flexibility and scoring functions in both ligand docking and virtual screening.展开更多
The long-term effectiveness of targeted cancer therapies is limited by the development of resistance.Although epigenetic reprogramming has been implicated in resistance,the mechanisms remain elusive.Herein,we demonstr...The long-term effectiveness of targeted cancer therapies is limited by the development of resistance.Although epigenetic reprogramming has been implicated in resistance,the mechanisms remain elusive.Herein,we demonstrate that increased chromatin accessibility is involved in adaptive BRAF inhibitor(BRAFi)-resistance in melanoma cells.We observed loss of chromatin assembly factor 1(CAF-1)and its related histone H3 lysine 9 trimethylation(H3K9me3)with adaptive BRAFi resistance.We further showed that depletion of CAF-1 provides chromatin plasticity for effective reprogramming by AP1 components to promote BRAFi resistance.Our data sug-gest that therapeutic approaches to restore H3K9me3 levels may compensate for the loss of CAF-1 and,in turn,suppress resistance to BRAF inhibitors.展开更多
AIM:To evaluate the current role of sorafenib,an oral multikinase inhibitor in the treatment of breast cancer.METHODS:An extensive search of the literature until March 2016 was carried out in Medline and clinicaltrial...AIM:To evaluate the current role of sorafenib,an oral multikinase inhibitor in the treatment of breast cancer.METHODS:An extensive search of the literature until March 2016 was carried out in Medline and clinicaltrials.gov,by using the search terms "sorafenib" and "breast cancer".Papers found were checked for further relevant publications.Overall,21 relevant studies were found,18 in advanced breast cancer(16 in stage Ⅳ and two in stages Ⅲ-Ⅳ) and three in early breast cancer.RESULTS:Among studies in advanced breast cancer,there were two trials with sorafenib as monotherapy,four trials of sorafenib in combination with taxanes,two in combination with capecitabine,one with gemcitabine and/or capecitabine,one with vinorelbine,one with bevacizumab,one with pemetrexed and one with ixabepilone,three trials of sorafenib in combination with endocrine therapy and two trials in women with brain metastases undergoing whole brain radiotherapy.In addition,there was one trial of sorafenib added to standard chemotherapy in the adjuvant setting,and two trials in the neoadjuvant setting.In general,sorafenib was well tolerated in breast cancer patients,though its dosage had to be adjusted in some trials,and discontinuation rates were high,particularly for the combination of sorafenib with anastrozole.Sorafenib monotherapy and combinations with taxanes,bevacizumab and ixabepilone showed inadequate efficacy,while efficacy results from combinations with gemcitabine and/or capecitabine and possibly tamoxifen were more promising.CONCLUSION:At present,sorafenib should not be used for the treatment of breast cancer outside of clinical trials and more clinical data are needed in order to support its standard use in breast cancer therapy.展开更多
文摘Colonic polyps may arise from BRAF inhibitor treatment of melanoma, possibly due to paradoxical activation of the mitogen-activated protein (MAP)-kinase pathway. In an alternative evidence based scenario, tubular colonic adenomas with APC gene mutations have also been identified in the context of BRAF inhibitor treatment, in the absence of mutations of MAPK genes. A minority of colorectal cancers develop by an alternative “serrated polyp pathway”. This article postulates a novel hypothesis, that the established phenotypic and molecular characteristics of serrated colonic polyps/CRC offer an intriguing insight into the pathobiology of BRAF inhibitor induced colonic polyps. Serrated polyps are characterized by a CpG island methylation phenotype, MLH1 silencing and cellular senescence. They also have BRAF mutations. The contention is that BRAF inhibitor induced polyps mimic the afore-described histology and molecular features of serrated polyps with the exception that instead of the presence of BRAF mutations they induce C-RAF homodimers and B-RAF: C-RAF heterodimers.
文摘Ameloblastoma is a benign but locally aggressive odontogenic neoplasm that accounts for 10% of all tumors arising in the mandible and maxilla (1). Eighty percent of ameloblastomas arise in the mandible, and they are usually found in young adults. It frequently recurs if not adequately
基金supported by grants from the National Natural Science Foundation of China(Grant No.81872290)the National Key R&D Program of China(Grant No.2017YFC0908500)
文摘BRAF is a serine/threonine kinase that harbors activating mutations in^7%of human malignancies and^60%of melanomas.Despite initial clinical responses to BRAF inhibitors,patients frequently develop drug resistance.To identify candidate therapeutic targets for BRAF inhibitor resistant melanoma,we conduct CRISPR screens in melanoma cells harboring an activating BRAF mutation that had also acquired resistance to BRAF inhibitors.To investigate the mechanisms and pathways enabling resistance to BRAF inhibitors in melanomas,we integrate expression,ATAC-seq,and CRISPR screen data.We identify the JUN family transcription factors and the ETS family transcription factor ETV5 as key regulators of CDK6,which together enable resistance to BRAF inhibitors in melanoma cells.Our findings reveal genes contributing to resistance to a selective BRAF inhibitor PLX4720,providing new insights into gene regulation in BRAF inhibitor resistant melanoma cells.
基金This work was supported by grants from the National Natural Science Foundation of China(No.81673917 and No.81973582)Guangdong Basic and Applied Basic Research Foundation(No.2019A1515110833)the Fundamental Research Funds of Shandong University(No.2019GN043).
文摘Objective:SOX4,a transcription factor,has been found to contribute to tumorigenesis in several cancers.This study was performed to determine whether SOX4 mediates BRAF inhibitor resistance in melanoma.Methods:Melanoma cell lines with acquired resistance to BRAF inhibitor(SK-MEL-5R,SK-MEL-28R,and A375R)were generated by adding escalating concentrations of PLX4032 into parental SK-MEL-5,SK-MEL-28,and A375 cells for>6 months.The expression of SOX4 and insulin-like growth factor 1 receptor(IGF-1R)was measured by quantitative real-time polymerase chain reaction(qRT-PCR)and Westem blotting.The downstream signaling of IGF-1R was detected by Westem blotting.SOX4 and IGF-1R overexpression or knockdown was conducted by lentivirus transfection.Cell viability and apoptosis were demonstrated by MTT and flow cytometry,respectively.The binding ability of SOX4 to IGF-1R promoter was determined by chromatin immunoprecipitation quantitative PCR assay.Results:SOX4 was upregulated in BRAF inhibitor-resistant melanoma cells as compared with parental cells(SK-MEL-5 group,1.02 vs.6.33;SK-MEL-28 group,1.03 vs.3.22;A375 group,1.00 vs.1.86;t=°7.069,29.26,and 5.291,respectively;all P<0.01),and PLX4032 treatment could not alter the expression of SOX4 in resistant cells.SOX4 overexpreesion attenuated the response of parental cells to PLX4032(for cell viability,SK-MEL-5 group:77.76%vs.104.28%,F=91.50;SK-MEL-28 group:60.59%vs.93.13%,F=171.8;A375 group:62.50%vs.80.87%,F=47.15.For apoptosis rates,SK-MEL-5 group:34.90%vs.14.31%,F=4.781;SK-MEL-28 group,40.8%vs.29.4%,F=13.32,P=0.063;A375 group:40.20%vs.17.09%,F=11.39;all P<0.05,otherwise indicated).While SOX4 knockdown enhanced the response of resistant cells to PLX4032(for cell viability,SK-MEL-5R group:93.75%vs.69.53%,F=94.45,SK-MEL-28R group:95.60%vs.66.79%,F=30.41,A375R group:95.51%vs.59.98%,F=111.6;for apoptosis rates,SK-MEL-5R group:16.2%vs.44.4%,F=25.67,SK-MEL-28R group:26.59%vs.44.20%,F=158.0,A375R group:5.98%vs.31.51%,F=14.35,and all P<0.01).Chromatin immunoprecipitation quantitative PCR assay demonstrated that SOX4 binded to the promoter of IGF-1R(1.04 vs.1.94[-1044 to-920 bp]and 0.110 vs.0.139[GAPDH],F=534.5,P<0.01).In addition,SOX4 overexpression increased IGF-1R end its downstream phosphorylated ERK,phosphorylated AKT,and phosphorylated STAT3 expression,while SOX4 knockdown exerted the opposite effects.Moreover,IGF-1R knockdown overcame SOX4 overexpreesion-induced PLX4032 resistance(cell viability:35.85%vs.52.79%vs.37.84%[A375 group,negative control group vs.SOX4 overexpressing group vs.SOX4 overexpressing+sh-IGF-1R group];apoptosis rates:25.30%vs.9.56%vs.22.26[A375 group,negative control group vs.SOX4 overexprassing group vs.SOX4 overexpressing+sh-IGF-1R group];F=13.01 and 41.87,respectively;all P<0.01),while IGF-1R overexpression abrogated SOX4 knockdown-induced response enhancement to PLX4032 for comparison of negative control group,sh-SOX4group and sh-SOX4+IGF-1 R overexpreesing group(cell viability:96.62%vs.86.86%vs.97.26%(A375R),98.15%vs.81.63%vs.98.49%[SK-MEL-5R];apoptosis rates:13.81%vs.32.00%vs.12.16[A375R],29.70%vs.41.40%vs.26.10%[SK-MEL-5R];F=13.56,12.86,38.81,and Conclusion:SOX4 mediates BRAF inhibitor resistance in melanoma through regulation of IGF-1R signaling.SOX4 might serve as a potential target for the treatment of BRAF inhibitor-resistant melanoma.39.85,respectively;all P<0.01).
文摘BACKGROUND Immune checkpoint inhibitors have revolutionized the treatment of patients with unresectable metastatic malignant melanoma.In addition to systemic side effects,several usually mild ocular adverse effects have been reported.We report a case of rarely reported vision-threatening bilateral panuveitis with serous retinal detachment,thickened choroid,and chorioretinal folds associated with dabrafenib and trametinib targeted therapy for B-Raf proto-oncogene serine/threonine kinase(BRAF)mutant metastatic cutaneous melanoma.CASE SUMMARY A 59-year-old female patient with metastatic melanoma treated with dabrafenib and trametinib presented with blurry vision and central scotoma lasting for 3 d in both eyes.Clinical examination and multimodal imaging revealed inflammatory cells in the anterior chamber,mild vitritis,bullous multiple serous retinal detachments,and chorioretinal folds in both eyes.Treatment with dabrafenib and trametinib was suspended,and the patient was treated with topical and intravenous corticosteroids followed by oral corticosteroid treatment with a tapering schedule.One and a half months after the disease onset,ocular morphological and functional improvement was noted.Due to the metastatic melanoma dissemination,BRAF/mitogen-activated protein kinase inhibitors were reintroduced and some mild ocular adverse effects reappeared,which later subsided after receiving oral corticosteroids.CONCLUSION Patients on combination therapy with dabrafenib and trametinib may rarely develop severe bilateral panuveitis with a good prognosis.Further studies have to establish potential usefulness of ophthalmological examination for asymptomatic patients.Furthermore,appropriate guidelines for managing panuveitis associated with dabrafenib and trametinib should be established.
基金supported by the National Natural Science Foundation of China(21102181,81302634 and 21572273)
文摘BRAF has been recognized as a promising target for cancer therapy. A number of crystal structures have been published. Molecular docking is one of the most effective techniques in the field of computer-aided drug design（CADD）. Appropriate protein conformation and docking method are essential for the successful virtual screening experiments. One approach considering protein flexibility and multiple docking methods was proposed in this study. Six DFG-in/αC-helix-out crystal structures of BRAF, three docking programs（Glide, GOLD and Ligand Fit） and 12 scoring functions were applied for the best combination by judging from the results of pose prediction and retrospective virtual screening（VS）. The most accurate results（mean RMSD of about 0.6 A） of pose prediction were obtained with two complex structures（PDB： 3 C4 C and 3 SKC） using Glide SP. From the retrospective VS, the most active compounds were identified by using the complex structure of 3 SKC, indicated by a ROC/AUC score of 0.998 and an EF of 20.6 at 5% of the database screen with Glide-SP. On the whole, PDB 3 SKC could achieve a higher rate of correct reproduction, a better enrichment and more diverse compounds. A comparison of 3 SKC and the other X-ray crystal structures led to a rationale for the docking results. PDB 3 SKC could achieve a broad range of sulfonamide substitutions through an expanded hydrophobic pocket formed by a further shift of the αC-helix. Our study emphasized the necessity and significance of protein flexibility and scoring functions in both ligand docking and virtual screening.
基金supported by the National Institutes of Health to R.B.D.(NIH5P30CA015083)the Mayo Clinic Cancer Center Ea-gles Cancer Fund(Z.W.)the Mayo Clinic Center for Biomedical Discovery(S.M.O).
文摘The long-term effectiveness of targeted cancer therapies is limited by the development of resistance.Although epigenetic reprogramming has been implicated in resistance,the mechanisms remain elusive.Herein,we demonstrate that increased chromatin accessibility is involved in adaptive BRAF inhibitor(BRAFi)-resistance in melanoma cells.We observed loss of chromatin assembly factor 1(CAF-1)and its related histone H3 lysine 9 trimethylation(H3K9me3)with adaptive BRAFi resistance.We further showed that depletion of CAF-1 provides chromatin plasticity for effective reprogramming by AP1 components to promote BRAFi resistance.Our data sug-gest that therapeutic approaches to restore H3K9me3 levels may compensate for the loss of CAF-1 and,in turn,suppress resistance to BRAF inhibitors.
文摘AIM:To evaluate the current role of sorafenib,an oral multikinase inhibitor in the treatment of breast cancer.METHODS:An extensive search of the literature until March 2016 was carried out in Medline and clinicaltrials.gov,by using the search terms "sorafenib" and "breast cancer".Papers found were checked for further relevant publications.Overall,21 relevant studies were found,18 in advanced breast cancer(16 in stage Ⅳ and two in stages Ⅲ-Ⅳ) and three in early breast cancer.RESULTS:Among studies in advanced breast cancer,there were two trials with sorafenib as monotherapy,four trials of sorafenib in combination with taxanes,two in combination with capecitabine,one with gemcitabine and/or capecitabine,one with vinorelbine,one with bevacizumab,one with pemetrexed and one with ixabepilone,three trials of sorafenib in combination with endocrine therapy and two trials in women with brain metastases undergoing whole brain radiotherapy.In addition,there was one trial of sorafenib added to standard chemotherapy in the adjuvant setting,and two trials in the neoadjuvant setting.In general,sorafenib was well tolerated in breast cancer patients,though its dosage had to be adjusted in some trials,and discontinuation rates were high,particularly for the combination of sorafenib with anastrozole.Sorafenib monotherapy and combinations with taxanes,bevacizumab and ixabepilone showed inadequate efficacy,while efficacy results from combinations with gemcitabine and/or capecitabine and possibly tamoxifen were more promising.CONCLUSION:At present,sorafenib should not be used for the treatment of breast cancer outside of clinical trials and more clinical data are needed in order to support its standard use in breast cancer therapy.
