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BACKGROUND: With poor prognosis and high mortality, pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal malignancies. Standard of care therapies for PDAC have included gemcitabine for the past three decades, although resistance often develops within weeks of chemotherapy initiation through an array of possible mechanisms. METHODS: We reanalyzed publicly available RNA-seq gene expression profiles of 28 PDAC patient-derived xenograft (PDX) models before and after a 21-day gemcitabine treatment using our validated analysis pipeline to identify molecular markers of intrinsic and acquired resistance. RESULTS: Using normalized RNA-seq quantification measurements, we first identified oxidative phosphorylation and interferon alpha pathways as the two most enriched cancer hallmark gene sets in the baseline gene expression profile associated with intrinsic gemcitabine resistance and sensitivity, respectively. Furthermore, we discovered strong correlations between drug-induced expression changes in glycolysis and oxidative phosphorylation genes and response to gemcitabine, which suggests that these pathways may be associated with acquired gemcitabine resistance mechanisms. Thus, we developed prediction models using baseline gene expression profiles in those pathways and validated them in another dataset of 12 PDAC models from Novartis. We also developed prediction models based on drug-induced expression changes in genes from the Molecular Signatures Database (MSigDB)'s curated 50 cancer hallmark gene sets. Finally, pathogenic TP53 mutations correlated with treatment resistance. CONCLUSION: Our results demonstrate that concurrent upregulation of both glycolysis and oxidative phosphorylation pathways occurs in vivo in PDAC PDXs following gemcitabine treatment and that pathogenic TP53 status had association with gemcitabine resistance in these models. Our findings may elucidate the molecular basis for gemcitabine resistance and provide insights for effective drug combination in PDAC chemotherapy.
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Desoxicitidina , Resistencia a Antineoplásicos , Gemcitabina , Neoplasias Pancreáticas , Proteína p53 Supresora de Tumor , Ensayos Antitumor por Modelo de Xenoinjerto , Desoxicitidina/análogos & derivados , Desoxicitidina/farmacología , Desoxicitidina/uso terapéutico , Humanos , Neoplasias Pancreáticas/tratamiento farmacológico , Neoplasias Pancreáticas/patología , Neoplasias Pancreáticas/genética , Resistencia a Antineoplásicos/genética , Proteína p53 Supresora de Tumor/metabolismo , Proteína p53 Supresora de Tumor/genética , Animales , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Carcinoma Ductal Pancreático/tratamiento farmacológico , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/patología , Carcinoma Ductal Pancreático/metabolismo , Ratones , Reprogramación MetabólicaRESUMEN
In 2005, several experts in tumor biomarker research publishe the REporting recommendations for Tumor MARKer prognostic studies (REMARK) criteria. Coupled with the subsequent Biospecimen Reporting for Improved Study Quality (BRISQ) criteria, these initiatives provide a framework for transparently reporting of the methods of study conduct and analyses.
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Biomarcadores de Tumor , Proyectos de Investigación , Humanos , PronósticoRESUMEN
BACKGROUND: In high-dimensional data (HDD) settings, the number of variables associated with each observation is very large. Prominent examples of HDD in biomedical research include omics data with a large number of variables such as many measurements across the genome, proteome, or metabolome, as well as electronic health records data that have large numbers of variables recorded for each patient. The statistical analysis of such data requires knowledge and experience, sometimes of complex methods adapted to the respective research questions. METHODS: Advances in statistical methodology and machine learning methods offer new opportunities for innovative analyses of HDD, but at the same time require a deeper understanding of some fundamental statistical concepts. Topic group TG9 "High-dimensional data" of the STRATOS (STRengthening Analytical Thinking for Observational Studies) initiative provides guidance for the analysis of observational studies, addressing particular statistical challenges and opportunities for the analysis of studies involving HDD. In this overview, we discuss key aspects of HDD analysis to provide a gentle introduction for non-statisticians and for classically trained statisticians with little experience specific to HDD. RESULTS: The paper is organized with respect to subtopics that are most relevant for the analysis of HDD, in particular initial data analysis, exploratory data analysis, multiple testing, and prediction. For each subtopic, main analytical goals in HDD settings are outlined. For each of these goals, basic explanations for some commonly used analysis methods are provided. Situations are identified where traditional statistical methods cannot, or should not, be used in the HDD setting, or where adequate analytic tools are still lacking. Many key references are provided. CONCLUSIONS: This review aims to provide a solid statistical foundation for researchers, including statisticians and non-statisticians, who are new to research with HDD or simply want to better evaluate and understand the results of HDD analyses.
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Investigación Biomédica , Objetivos , Humanos , Proyectos de InvestigaciónRESUMEN
An important element of precision medicine is the ability to identify, for a specific therapy, those patients for whom benefits of that therapy meaningfully exceed the risks. To achieve this goal, treatment effect usually is examined across subgroups defined by a variety of factors, including demographic, clinical, or pathologic characteristics or by molecular attributes of patients or their disease. Frequently such subgroups are defined by the measurement of biomarkers. Even though such examination is necessary when pursuing this goal, the evaluation of treatment effect across a variety of subgroups is statistically fraught due to both the danger of inflated false-positive error rate from multiple testing and the inherent insensitivity to how treatment effects differ across subgroups.Pre-specification of subgroup analyses with appropriate control of false-positive (i.e. type I) error is recommended when possible. However, when subgroups are specified by biomarkers, which could be measured by different assays and might lack established interpretation criteria, such as cut-offs, it might not be possible to fully specify those subgroups at the time a new therapy is ready for definitive evaluation in a Phase 3 trial. In these situations, further refinement and evaluation of treatment effect in biomarker-defined subgroups might have to take place within the trial. A common scenario is that evidence suggests that treatment effect is a monotone function of a biomarker value, but optimal cut-offs for therapy decisions are not known. In this setting, hierarchical testing strategies are widely used, where testing is first conducted in a particular biomarker-positive subgroup and then is conducted in the expanded pool of biomarker-positive and biomarker-negative patients, with control for multiple testing. A serious limitation of this approach is the logical inconsistency of excluding the biomarker-negatives when evaluating effects in the biomarker-positives, yet allowing the biomarker-positives to drive the assessment of whether a conclusion of benefit could be extrapolated to the biomarker-negative subgroup.Examples from oncology and cardiology are described to illustrate the challenges and pitfalls. Recommendations are provided for statistically valid and logically consistent subgroup testing in these scenarios as alternatives to reliance on hierarchical testing alone, and approaches for exploratory assessment of continuous biomarkers as treatment effect modifiers are discussed.
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Medicina de Precisión , Humanos , BiomarcadoresRESUMEN
BACKGROUND: Participation of adolescents and young adults (AYAs) in oncology clinical trials is important to ensure adequate opportunities for AYA patients to contribute to, and benefit from, advances in cancer treatment. METHODS: Accrual data for National Cancer Institute (NCI) Cancer Therapy Evaluation Program (CTEP) cooperative group-led treatment trials were examined to assess enrollment of newly diagnosed AYA patients (15-39 years) during the period 2004-2019, with particular interest in comparing enrollment before launch of the NCI National Clinical Trials Network (NCTN) to after. All phase 2, 2/3, and 3 trials activated during the period between January 1, 2004, and December 31, 2019, were identified (n = 1568) and reduced to a set of 304 that met predetermined criteria to focus on cooperative group-led trials that involved therapy for newly diagnosed cancer and had age eligibility overlapping the AYA range. The proportion of AYA patients relative to total accrual, along with 95% bootstrapped CI was calculated for patients enrolled pre-NCTN and post-NCTN. RESULTS: AYA accrual comprised 9.5% (95% CI, 7.6-11.8) pre-NCTN compared with 14.0% (95% CI, 9.9-18.3) post-NCTN. The mean difference in proportions post-NCTN compared with pre-NCTN was 4.4% (0.7%-8.3%). CONCLUSIONS: These results indicate an increase in AYA participation in trials conducted within the NCTN relative to the pre-NCTN period. This suggests an awareness and utilization of NCTN trials for AYAs with cancer.
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Oncología Médica , Neoplasias , Academias e Institutos , Adolescente , Recolección de Datos , Humanos , National Cancer Institute (U.S.) , Neoplasias/terapia , Estados Unidos , Adulto JovenRESUMEN
BACKGROUND: Indian natural products have been anecdotally used for cancer treatment but with limited efficacy. To better understand their mechanism, we examined the publicly available data for the activity of Indian natural products in the NCI-60 cell line panel. METHODS: We examined associations of molecular genomic features in the well-characterized NCI-60 cancer cell line panel with in vitro response to treatment with 75 compounds derived from Indian plant-based natural products. We analyzed expression measures for annotated transcripts, lncRNAs, and miRNAs, and protein-changing single nucleotide variants in cancer-related genes. We also examined the similarities between cancer cell line response to Indian natural products and response to reference anti-tumor compounds recorded in a U.S. National Cancer Institute (NCI) Developmental Therapeutics Program database. RESULTS: Hierarchical clustering based on cell line response measures identified clustering of Phyllanthus and cucurbitacin products with known anti-tumor agents with anti-mitotic mechanisms of action. Curcumin and curcuminoids mostly clustered together. We found associations of response to Indian natural products with expression of multiple genes, notably including SLC7A11 involved in solute transport and ATAD3A and ATAD3B encoding mitochondrial ATPase proteins, as well as significant associations with functional single nucleotide variants, including BRAF V600E. CONCLUSION: These findings suggest potential mechanisms of action and novel associations of in vitro response with gene expression and some cancer-related mutations that increase our understanding of these Indian natural products.
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Antineoplásicos , Productos Biológicos , Neoplasias , ATPasas Asociadas con Actividades Celulares Diversas , Antineoplásicos/farmacología , Productos Biológicos/farmacología , Línea Celular Tumoral , Humanos , Proteínas de la Membrana , Proteínas Mitocondriales , National Cancer Institute (U.S.) , Neoplasias/tratamiento farmacológico , Neoplasias/genética , Nucleótidos , Farmacogenética , Estados UnidosRESUMEN
Treatment selection biomarkers are those that can be useful in guiding choice of therapy. Just as new therapies require evaluation in appropriately designed clinical trials to determine their benefit, therapy selection biomarkers require evaluation in appropriately designed studies. These studies may be prospective clinical trials or retrospective studies based on specimens stored from a completed clinical trial. Ideally, patient treatment assignments should be randomized, and consideration should be given to an appropriate sample size-either for prospective planning of a new study or access to a sufficient number of stored specimens. Here, we develop a novel sample size method for estimation of a confidence interval of specified average width, for an intuitively appealing previously proposed parameter that reflects the expected benefit of using biomarker-guided therapy relative to a standard-of-care therapy. The estimation approach combines Monte Carlo and regression to result in a procedure that performs well over a range of scenarios. Although derived under a specific Cox proportional hazards regression model, robustness to model violations is demonstrated by evaluation under accelerated failure time and cure models. The sample size method produces adequate or conservative sample size estimates under a range of scenarios. Computer code in R and C++, and applications for Mac and Windows are made available for implementation of the sample size estimation procedure. The method is applied to a real data setting and results discussed.
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Tamaño de la Muestra , Biomarcadores , Humanos , Modelos de Riesgos Proporcionales , Estudios Prospectivos , Estudios RetrospectivosRESUMEN
BACKGROUND: In order to correctly decode phenotypic information from RNA-sequencing (RNA-seq) data, careful selection of the RNA-seq quantification measure is critical for inter-sample comparisons and for downstream analyses, such as differential gene expression between two or more conditions. Several methods have been proposed and continue to be used. However, a consensus has not been reached regarding the best gene expression quantification method for RNA-seq data analysis. METHODS: In the present study, we used replicate samples from each of 20 patient-derived xenograft (PDX) models spanning 15 tumor types, for a total of 61 human tumor xenograft samples available through the NCI patient-derived model repository (PDMR). We compared the reproducibility across replicate samples based on TPM (transcripts per million), FPKM (fragments per kilobase of transcript per million fragments mapped), and normalized counts using coefficient of variation, intraclass correlation coefficient, and cluster analysis. RESULTS: Our results revealed that hierarchical clustering on normalized count data tended to group replicate samples from the same PDX model together more accurately than TPM and FPKM data. Furthermore, normalized count data were observed to have the lowest median coefficient of variation (CV), and highest intraclass correlation (ICC) values across all replicate samples from the same model and for the same gene across all PDX models compared to TPM and FPKM data. CONCLUSION: We provided compelling evidence for a preferred quantification measure to conduct downstream analyses of PDX RNA-seq data. To our knowledge, this is the first comparative study of RNA-seq data quantification measures conducted on PDX models, which are known to be inherently more variable than cell line models. Our findings are consistent with what others have shown for human tumors and cell lines and add further support to the thesis that normalized counts are the best choice for the analysis of RNA-seq data across samples.
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Secuenciación de Nucleótidos de Alto Rendimiento , ARN , Perfilación de la Expresión Génica , Humanos , RNA-Seq , Reproducibilidad de los Resultados , Análisis de Secuencia de ARNRESUMEN
Liquid biopsy, particularly the analysis of circulating tumor DNA (ctDNA), has demonstrated considerable promise for numerous clinical intended uses. Successful validation and commercialization of novel ctDNA tests have the potential to improve the outcomes of patients with cancer. The goal of the Blood Profiling Atlas Consortium (BloodPAC) is to accelerate the development and validation of liquid biopsy assays that will be introduced into the clinic. To accomplish this goal, the BloodPAC conducts research in the following areas: Data Collection and Analysis within the BloodPAC Data Commons; Preanalytical Variables; Analytical Variables; Patient Context Variables; and Reimbursement. In this document, the BloodPAC's Analytical Variables Working Group (AV WG) attempts to define a set of generic analytical validation protocols tailored for ctDNA-based Next-Generation Sequencing (NGS) assays. Analytical validation of ctDNA assays poses several unique challenges that primarily arise from the fact that very few tumor-derived DNA molecules may be present in circulation relative to the amount of nontumor-derived cell-free DNA (cfDNA). These challenges include the exquisite level of sensitivity and specificity needed to detect ctDNA, the potential for false negatives in detecting these rare molecules, and the increased reliance on contrived samples to attain sufficient ctDNA for analytical validation. By addressing these unique challenges, the BloodPAC hopes to expedite sponsors' presubmission discussions with the Food and Drug Administration (FDA) with the protocols presented herein. By sharing best practices with the broader community, this work may also save the time and capacity of FDA reviewers through increased efficiency.
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Biomarcadores de Tumor/sangre , ADN Tumoral Circulante/sangre , Guías como Asunto , Secuenciación de Nucleótidos de Alto Rendimiento/normas , Humanos , Biopsia Líquida , Neoplasias/sangre , Neoplasias/patología , Estándares de Referencia , Estudios de Validación como AsuntoRESUMEN
The nuclear proliferation biomarker Ki67 has potential prognostic, predictive, and monitoring roles in breast cancer. Unacceptable between-laboratory variability has limited its clinical value. The International Ki67 in Breast Cancer Working Group investigated whether Ki67 immunohistochemistry can be analytically validated and standardized across laboratories using automated machine-based scoring. Sets of pre-stained core-cut biopsy sections of 30 breast tumors were circulated to 14 laboratories for scanning and automated assessment of the average and maximum percentage of tumor cells positive for Ki67. Seven unique scanners and 10 software platforms were involved in this study. Pre-specified analyses included evaluation of reproducibility between all laboratories (primary) as well as among those using scanners from a single vendor (secondary). The primary reproducibility metric was intraclass correlation coefficient between laboratories, with success considered to be intraclass correlation coefficient >0.80. Intraclass correlation coefficient for automated average scores across 16 operators was 0.83 (95% credible interval: 0.73-0.91) and intraclass correlation coefficient for maximum scores across 10 operators was 0.63 (95% credible interval: 0.44-0.80). For the laboratories using scanners from a single vendor (8 score sets), intraclass correlation coefficient for average automated scores was 0.89 (95% credible interval: 0.81-0.96), which was similar to the intraclass correlation coefficient of 0.87 (95% credible interval: 0.81-0.93) achieved using these same slides in a prior visual-reading reproducibility study. Automated machine assessment of average Ki67 has the potential to achieve between-laboratory reproducibility similar to that for a rigorously standardized pathologist-based visual assessment of Ki67. The observed intraclass correlation coefficient was worse for maximum compared to average scoring methods, suggesting that maximum score methods may be suboptimal for consistent measurement of proliferation. Automated average scoring methods show promise for assessment of Ki67 scoring, but requires further standardization and subsequent clinical validation.
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Biomarcadores de Tumor/análisis , Neoplasias de la Mama/patología , Procesamiento de Imagen Asistido por Computador/normas , Inmunohistoquímica/normas , Antígeno Ki-67/análisis , Femenino , Humanos , Inmunohistoquímica/métodos , Reproducibilidad de los ResultadosRESUMEN
AIMS: The nuclear proliferation marker Ki67 assayed by immunohistochemistry has multiple potential uses in breast cancer, but an unacceptable level of interlaboratory variability has hampered its clinical utility. The International Ki67 in Breast Cancer Working Group has undertaken a systematic programme to determine whether Ki67 measurement can be analytically validated and standardised among laboratories. This study addresses whether acceptable scoring reproducibility can be achieved on excision whole sections. METHODS AND RESULTS: Adjacent sections from 30 primary ER+ breast cancers were centrally stained for Ki67 and sections were circulated among 23 pathologists in 12 countries. All pathologists scored Ki67 by two methods: (i) global: four fields of 100 tumour cells each were selected to reflect observed heterogeneity in nuclear staining; (ii) hot-spot: the field with highest apparent Ki67 index was selected and up to 500 cells scored. The intraclass correlation coefficient (ICC) for the global method [confidence interval (CI) = 0.87; 95% CI = 0.799-0.93] marginally met the prespecified success criterion (lower 95% CI ≥ 0.8), while the ICC for the hot-spot method (0.83; 95% CI = 0.74-0.90) did not. Visually, interobserver concordance in location of selected hot-spots varies between cases. The median times for scoring were 9 and 6 min for global and hot-spot methods, respectively. CONCLUSIONS: The global scoring method demonstrates adequate reproducibility to warrant next steps towards evaluation for technical and clinical validity in appropriate cohorts of cases. The time taken for scoring by either method is practical using counting software we are making publicly available. Establishment of external quality assessment schemes is likely to improve the reproducibility between laboratories further.
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Biomarcadores de Tumor/análisis , Neoplasias de la Mama , Inmunohistoquímica/normas , Antígeno Ki-67/análisis , Patología Clínica/normas , Femenino , Humanos , Variaciones Dependientes del Observador , Reproducibilidad de los ResultadosRESUMEN
The US National Cancer Institute (NCI), in collaboration with scientists representing multiple areas of expertise relevant to 'omics'-based test development, has developed a checklist of criteria that can be used to determine the readiness of omics-based tests for guiding patient care in clinical trials. The checklist criteria cover issues relating to specimens, assays, mathematical modelling, clinical trial design, and ethical, legal and regulatory aspects. Funding bodies and journals are encouraged to consider the checklist, which they may find useful for assessing study quality and evidence strength. The checklist will be used to evaluate proposals for NCI-sponsored clinical trials in which omics tests will be used to guide therapy.
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Ensayos Clínicos como Asunto/métodos , Genómica , Proyectos de Investigación , Lista de Verificación , Ensayos Clínicos como Asunto/economía , Ensayos Clínicos como Asunto/ética , Ensayos Clínicos como Asunto/normas , Estudios de Evaluación como Asunto , Genómica/ética , Humanos , Modelos Biológicos , National Cancer Institute (U.S.)/economía , Medicina de Precisión/ética , Medicina de Precisión/métodos , Medicina de Precisión/normas , Proyectos de Investigación/normas , Manejo de Especímenes , Estados UnidosRESUMEN
BACKGROUND/AIMS: Biomarker-stratified outcome-adaptive randomization trials, in which randomization probabilities depend on both biomarker value and outcomes of previously treated patients, are receiving increased attention in oncology research. Data from these trials can also form the basis of investigation of additional biomarkers that may not have been incorporated into the original trial design. In this article, we investigate the validity of a standard analytical method that utilizes data from a biomarker-stratified outcome-adaptive randomization trial to assess the effect of a newly identified biomarker on patient outcomes. METHODS: In the context of an ancillary biomarker study for a two-arm phase II trial with a response endpoint, we conduct analytic and simulation studies to investigate bias in estimated biomarker effects under outcome-adaptive randomization. Conditions under which bias arises and magnitude of the bias are examined in several settings. We then propose unbiased estimators of biomarker effects with appropriate variance estimators. RESULTS: We demonstrate that use of biomarker-stratified outcome-adaptive randomization perturbs the patient population and treatment assignments. Consequently, application of standard analysis methods to data from an outcome-adaptive randomization trial either to estimate prognostic effect of a new biomarker in uniformly treated patients or to estimate effect of treatment in relation to the new biomarker can lead to substantially biased estimates. The proposed adjusted estimators are asymptotically unbiased, and the proposed variance estimators correctly reflect the sample variability in the estimators. CONCLUSION: This article demonstrates existence of bias when standard, naïve statistical methods are utilized to assess biomarker effects using data from a biomarker-stratified outcome-adaptive randomization trial, and hence that results from naïve analyses must be interpreted with great caution. These findings highlight that, in an era where data and specimens are increasingly being shared for biomarker studies, care must be taken to document and understand implications of the study design under which specimens or data have been obtained.
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Sesgo , Biomarcadores , Modelos Estadísticos , Ensayos Clínicos Controlados Aleatorios como Asunto/métodos , Ensayos Clínicos Fase II como Asunto , Humanos , Oportunidad Relativa , Probabilidad , Ensayos Clínicos Controlados Aleatorios como Asunto/estadística & datos numéricos , Proyectos de Investigación , Estudios Retrospectivos , Tamaño de la MuestraRESUMEN
Omics technologies that generate a large amount of molecular data characterizing biospecimens have the potential to provide information about patients' disease characteristics above and beyond standard clinical features. By combining information from a large number of features into a multivariable model, called a biomarker signature, there is the opportunity to identify distinct subgroups of patients for whom treatment decisions can be personalized. The key challenge is to derive a signature with good performance and appropriately characterize its performance. We summarize the key statistical issues and methods for developing and validating biomarker signatures, using examples from the literature for illustration.
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Biomarcadores/análisis , Toma de Decisiones Clínicas , Modelos Estadísticos , Humanos , Análisis MultivarianteRESUMEN
Although investigators in the imaging community have been active in developing and evaluating quantitative imaging biomarkers (QIBs), the development and implementation of QIBs have been hampered by the inconsistent or incorrect use of terminology or methods for technical performance and statistical concepts. Technical performance is an assessment of how a test performs in reference objects or subjects under controlled conditions. In this article, some of the relevant statistical concepts are reviewed, methods that can be used for evaluating and comparing QIBs are described, and some of the technical performance issues related to imaging biomarkers are discussed. More consistent and correct use of terminology and study design principles will improve clinical research, advance regulatory science, and foster better care for patients who undergo imaging studies.
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Biomarcadores/análisis , Diagnóstico por Imagen/métodos , Sesgo , Fantasmas de Imagen , Valores de Referencia , Reproducibilidad de los Resultados , Terminología como AsuntoRESUMEN
Although an important biomarker in breast cancer, Ki67 lacks scoring standardization, which has limited its clinical use. Our previous study found variability when laboratories used their own scoring methods on centrally stained tissue microarray slides. In this current study, 16 laboratories from eight countries calibrated to a specific Ki67 scoring method and then scored 50 centrally MIB-1 stained tissue microarray cases. Simple instructions prescribed scoring pattern and staining thresholds for determination of the percentage of stained tumor cells. To calibrate, laboratories scored 18 'training' and 'test' web-based images. Software tracked object selection and scoring. Success for the calibration was prespecified as Root Mean Square Error of scores compared with reference <0.6 and Maximum Absolute Deviation from reference <1.0 (log2-transformed data). Prespecified success criteria for tissue microarray scoring required intraclass correlation significantly >0.70 but aiming for observed intraclass correlation ≥0.90. Laboratory performance showed non-significant but promising trends of improvement through the calibration exercise (mean Root Mean Square Error decreased from 0.6 to 0.4, Maximum Absolute Deviation from 1.6 to 0.9; paired t-test: P=0.07 for Root Mean Square Error, 0.06 for Maximum Absolute Deviation). For tissue microarray scoring, the intraclass correlation estimate was 0.94 (95% credible interval: 0.90-0.97), markedly and significantly >0.70, the prespecified minimum target for success. Some discrepancies persisted, including around clinically relevant cutoffs. After calibrating to a common scoring method via a web-based tool, laboratories can achieve high inter-laboratory reproducibility in Ki67 scoring on centrally stained tissue microarray slides. Although these data are potentially encouraging, suggesting that it may be possible to standardize scoring of Ki67 among pathology laboratories, clinically important discrepancies persist. Before this biomarker could be recommended for clinical use, future research will need to extend this approach to biopsies and whole sections, account for staining variability, and link to outcomes.
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Biomarcadores de Tumor/análisis , Neoplasias de la Mama/patología , Inmunohistoquímica/normas , Antígeno Ki-67/análisis , Análisis de Matrices Tisulares/normas , Femenino , HumanosRESUMEN
BACKGROUND: The intraclass correlation coefficient (ICC) is widely used in biomedical research to assess the reproducibility of measurements between raters, labs, technicians, or devices. For example, in an inter-rater reliability study, a high ICC value means that noise variability (between-raters and within-raters) is small relative to variability from patient to patient. A confidence interval or Bayesian credible interval for the ICC is a commonly reported summary. Such intervals can be constructed employing either frequentist or Bayesian methodologies. METHODS: This study examines the performance of three different methods for constructing an interval in a two-way, crossed, random effects model without interaction: the Generalized Confidence Interval method (GCI), the Modified Large Sample method (MLS), and a Bayesian method based on a noninformative prior distribution (NIB). Guidance is provided on interval construction method selection based on study design, sample size, and normality of the data. We compare the coverage probabilities and widths of the different interval methods. RESULTS: We show that, for the two-way, crossed, random effects model without interaction, care is needed in interval method selection because the interval estimates do not always have properties that the user expects. While different methods generally perform well when there are a large number of levels of each factor, large differences between the methods emerge when the number of one or more factors is limited. In addition, all methods are shown to lack robustness to certain hard-to-detect violations of normality when the sample size is limited. CONCLUSIONS: Decision rules and software programs for interval construction are provided for practical implementation in the two-way, crossed, random effects model without interaction. All interval methods perform similarly when the data are normal and there are sufficient numbers of levels of each factor. The MLS and GCI methods outperform the NIB when one of the factors has a limited number of levels and the data are normally distributed or nearly normally distributed. None of the methods work well if the number of levels of a factor are limited and data are markedly non-normal. The software programs are implemented in the popular R language.
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Interpretación Estadística de Datos , Modelos Estadísticos , Proyectos de Investigación , Análisis de Varianza , Teorema de Bayes , Intervalos de Confianza , Humanos , Reproducibilidad de los Resultados , Programas InformáticosRESUMEN
Histone deacetylases (HDACs) and sirtuins (SIRTs) are important epigenetic regulators of cancer pathways. There is a limited understanding of how transcriptional regulation of their genes is affected by chemotherapeutic agents, and how such transcriptional changes affect tumour sensitivity to drug treatment. We investigated the concerted transcriptional response of HDAC and SIRT genes to 15 approved antitumor agents in the NCI-60 cancer cell line panel. Antitumor agents with diverse mechanisms of action induced upregulation or downregulation of multiple HDAC and SIRT genes. HDAC5 was upregulated by dasatinib and erlotinib in the majority of the cell lines. Tumour cell line sensitivity to kinase inhibitors was associated with upregulation of HDAC5, HDAC1, and several SIRT genes. We confirmed changes in HDAC and SIRT expression in independent datasets. We also experimentally validated the upregulation of HDAC5 mRNA and protein expression by dasatinib in the highly sensitive IGROV1 cell line. HDAC5 was not upregulated in the UACC-257 cell line resistant to dasatinib. The effects of cancer drug treatment on expression of HDAC and SIRT genes may influence chemosensitivity and may need to be considered during chemotherapy.
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Antineoplásicos , Neoplasias , Sirtuinas , Dasatinib/farmacología , Metilación de ADN , Línea Celular Tumoral , Sirtuinas/genética , Sirtuinas/metabolismo , Antineoplásicos/farmacología , Inhibidores de Histona Desacetilasas/farmacología , Neoplasias/tratamiento farmacológico , Neoplasias/genéticaRESUMEN
PURPOSE: Subprotocol K2 (EAY131-K2) of the NCI-MATCH platform trial was an open-label, single-arm, phase II study designed to evaluate the antitumor efficacy of the oral FGFR1-4 inhibitor, erdafitinib, in patients with tumors harboring FGFR1-4 mutations or fusions. METHODS: Central confirmation of tumor FGFR1-4 mutations or fusions was required for outcome analysis. Patients with urothelial carcinoma were excluded. Enrolled subjects received oral erdafitinib at a starting dose of 8 mg daily continuously until intolerable toxicity or disease progression. The primary end point was objective response rate (ORR) with key secondary end points of safety, progression-free survival (PFS), and overall survival (OS). RESULTS: Thirty-five patients were enrolled, and 25 patients were included in the primary efficacy analysis as prespecified in the protocol. The median age was 61 years, and 52% of subjects had received ≥3 previous lines of therapy. The confirmed ORR was 16% (4 of 25 [90% CI, 5.7 to 33.0], P = .034 against the null rate of 5%). An additional seven patients experienced stable disease as best-confirmed response. Four patients had a prolonged PFS including two with recurrent WHO grade IV, IDH1-/2-wildtype glioblastoma. The median PFS and OS were 3.6 months and 11.0 months, respectively. Erdafitinib was manageable with no new safety signals. CONCLUSION: This study met its primary end point in patients with several pretreated solid tumor types harboring FGFR1-3 mutations or fusions. These findings support advancement of erdafitinib for patients with fibroblast growth factor receptor-altered tumors outside of currently approved indications in a potentially tumor-agnostic manner.