GATA2 regulates the erythropoietin receptor in t(12;21) ALL.

The t(12;21) (p13;q22) chromosomal translocation resulting in the ETV6/RUNX1 fusion gene is the most frequent structural cytogenetic abnormality in children with acute lymphoblastic leukemia (ALL). The erythropoietin receptor (EPOR), usually associated with erythroid progenitor cells, is highly expressed in ETV6/RUNX1 positive cases compared to other B-lineage ALL subtypes. Gene expression analysis of a microarray database and direct quantitative analysis of patient samples revealed strong correlation between EPOR and GATA2 expression in ALL, and higher expression of GATA2 in t(12;21) patients. The mechanism of EPOR regulation was mainly investigated using two B-ALL cell lines: REH, which harbor and express the ETV6/RUNX1 fusion gene; and NALM-6, which do not. Expression of EPOR was increased in REH cells compared to NALM-6 cells. Moreover, of the six GATA family members only GATA2 was differentially expressed with substantially higher levels present in REH cells. GATA2 was shown to bind to the EPOR 5'-UTR in REH, but did not bind in NALM-6 cells. Overexpression of GATA2 led to an increase in EPOR expression in REH cells only, indicating that GATA2 regulates EPOR but is dependent on the cellular context. Both EPOR and GATA2 are hypomethylated and associated with increased mRNA expression in REH compared to NALM-6 cells. Decitabine treatment effectively reduced methylation of CpG sites in the GATA2 promoter leading to increased GATA2 expression in both cell lines. Although Decitabine also reduced an already low level of methylation of the EPOR in NALM-6 cells there was no increase in EPOR expression. Furthermore, EPOR and GATA2 are regulated post-transcriptionally by miR-362 and miR-650, respectively. Overall our data show that EPOR expression in t(12;21) B-ALL cells, is regulated by GATA2 and is mediated through epigenetic, transcriptional and post-transcriptional mechanisms, contingent upon the genetic subtype of the disease

An integrative workflow to study large-scale biochemical networks

I propose an integrative workflow to study large-scale biochemical networks by combining omics data, network structure and dynamical analysis to unravel disease mechanisms. Using the workflow, I identified core regulatory networks from the E2F1 network underlying EMT in bladder and breast cancer and detected disease signatures and drug targets, which were experimentally validated. Further, I developed a hybrid modeling framework that combines ODE- with logical-models to analyze the dynamics of large-scale non-linear systems. This thesis is a contribution to interdisciplinary cancer research

Otimização multi-objetivo em aprendizado de máquina

Orientador: Fernando José Von ZubenTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Elétrica e de ComputaçãoResumo: Regressão logística multinomial regularizada, classificação multi-rótulo e aprendizado multi-tarefa são exemplos de problemas de aprendizado de máquina em que objetivos conflitantes, como funções de perda e penalidades que promovem regularização, devem ser simultaneamente minimizadas. Portanto, a perspectiva simplista de procurar o modelo de aprendizado com o melhor desempenho deve ser substituída pela proposição e subsequente exploração de múltiplos modelos de aprendizado eficientes, cada um caracterizado por um compromisso (trade-off) distinto entre os objetivos conflitantes. Comitês de máquinas e preferências a posteriori do tomador de decisão podem ser implementadas visando explorar adequadamente este conjunto diverso de modelos de aprendizado eficientes, em busca de melhoria de desempenho. A estrutura conceitual multi-objetivo para aprendizado de máquina é suportada por três etapas: (1) Modelagem multi-objetivo de cada problema de aprendizado, destacando explicitamente os objetivos conflitantes envolvidos; (2) Dada a formulação multi-objetivo do problema de aprendizado, por exemplo, considerando funções de perda e termos de penalização como objetivos conflitantes, soluções eficientes e bem distribuídas ao longo da fronteira de Pareto são obtidas por um solver determinístico e exato denominado NISE (do inglês Non-Inferior Set Estimation); (3) Esses modelos de aprendizado eficientes são então submetidos a um processo de seleção de modelos que opera com preferências a posteriori, ou a filtragem e agregação para a síntese de ensembles. Como o NISE é restrito a problemas de dois objetivos, uma extensão do NISE capaz de lidar com mais de dois objetivos, denominada MONISE (do inglês Many-Objective NISE), também é proposta aqui, sendo uma contribuição adicional que expande a aplicabilidade da estrutura conceitual proposta. Para atestar adequadamente o mérito da nossa abordagem multi-objetivo, foram realizadas investigações mais específicas, restritas à aprendizagem de modelos lineares regularizados: (1) Qual é o mérito relativo da seleção a posteriori de um único modelo de aprendizado, entre os produzidos pela nossa proposta, quando comparado com outras abordagens de modelo único na literatura? (2) O nível de diversidade dos modelos de aprendizado produzidos pela nossa proposta é superior àquele alcançado por abordagens alternativas dedicadas à geração de múltiplos modelos de aprendizado? (3) E quanto à qualidade de predição da filtragem e agregação dos modelos de aprendizado produzidos pela nossa proposta quando aplicados a: (i) classificação multi-classe, (ii) classificação desbalanceada, (iii) classificação multi-rótulo, (iv) aprendizado multi-tarefa, (v) aprendizado com multiplos conjuntos de atributos? A natureza determinística de NISE e MONISE, sua capacidade de lidar adequadamente com a forma da fronteira de Pareto em cada problema de aprendizado, e a garantia de sempre obter modelos de aprendizado eficientes são aqui pleiteados como responsáveis pelos resultados promissores alcançados em todas essas três frentes de investigação específicasAbstract: Regularized multinomial logistic regression, multi-label classification, and multi-task learning are examples of machine learning problems in which conflicting objectives, such as losses and regularization penalties, should be simultaneously minimized. Therefore, the narrow perspective of looking for the learning model with the best performance should be replaced by the proposition and further exploration of multiple efficient learning models, each one characterized by a distinct trade-off among the conflicting objectives. Committee machines and a posteriori preferences of the decision-maker may be implemented to properly explore this diverse set of efficient learning models toward performance improvement. The whole multi-objective framework for machine learning is supported by three stages: (1) The multi-objective modelling of each learning problem, explicitly highlighting the conflicting objectives involved; (2) Given the multi-objective formulation of the learning problem, for instance, considering loss functions and penalty terms as conflicting objective functions, efficient solutions well-distributed along the Pareto front are obtained by a deterministic and exact solver named NISE (Non-Inferior Set Estimation); (3) Those efficient learning models are then subject to a posteriori model selection, or to ensemble filtering and aggregation. Given that NISE is restricted to two objective functions, an extension for many objectives, named MONISE (Many Objective NISE), is also proposed here, being an additional contribution and expanding the applicability of the proposed framework. To properly access the merit of our multi-objective approach, more specific investigations were conducted, restricted to regularized linear learning models: (1) What is the relative merit of the a posteriori selection of a single learning model, among the ones produced by our proposal, when compared with other single-model approaches in the literature? (2) Is the diversity level of the learning models produced by our proposal higher than the diversity level achieved by alternative approaches devoted to generating multiple learning models? (3) What about the prediction quality of ensemble filtering and aggregation of the learning models produced by our proposal on: (i) multi-class classification, (ii) unbalanced classification, (iii) multi-label classification, (iv) multi-task learning, (v) multi-view learning? The deterministic nature of NISE and MONISE, their ability to properly deal with the shape of the Pareto front in each learning problem, and the guarantee of always obtaining efficient learning models are advocated here as being responsible for the promising results achieved in all those three specific investigationsDoutoradoEngenharia de ComputaçãoDoutor em Engenharia Elétrica2014/13533-0FAPES

Evolutionary approaches for feature selection in biological data

Data mining techniques have been used widely in many areas such as business, science, engineering and medicine. The techniques allow a vast amount of data to be explored in order to extract useful information from the data. One of the foci in the health area is finding interesting biomarkers from biomedical data. Mass throughput data generated from microarrays and mass spectrometry from biological samples are high dimensional and is small in sample size. Examples include DNA microarray datasets with up to 500,000 genes and mass spectrometry data with 300,000 m/z values. While the availability of such datasets can aid in the development of techniques/drugs to improve diagnosis and treatment of diseases, a major challenge involves its analysis to extract useful and meaningful information. The aims of this project are: 1) to investigate and develop feature selection algorithms that incorporate various evolutionary strategies, 2) using the developed algorithms to find the “most relevant” biomarkers contained in biological datasets and 3) and evaluate the goodness of extracted feature subsets for relevance (examined in terms of existing biomedical domain knowledge and from classification accuracy obtained using different classifiers). The project aims to generate good predictive models for classifying diseased samples from control

Cutaneous Melanoma Classification: The Importance of High-Throughput Genomic Technologies

Cutaneous melanoma is an aggressive tumor responsible for 90% of mortality related to skin cancer. In the recent years, the discovery of driving mutations in melanoma has led to better treatment approaches. The last decade has seen a genomic revolution in the field of cancer. Such genomic revolution has led to the production of an unprecedented mole of data. High-throughput genomic technologies have facilitated the genomic, transcriptomic and epigenomic profiling of several cancers, including melanoma. Nevertheless, there are a number of newer genomic technologies that have not yet been employed in large studies. In this article we describe the current classification of cutaneous melanoma, we review the current knowledge of the main genetic alterations of cutaneous melanoma and their related impact on targeted therapies, and we describe the most recent highthroughput genomic technologies, highlighting their advantages and disadvantages. We hope that the current review will also help scientists to identify the most suitable technology to address melanoma-related relevant questions. The translation of this knowledge and all actual advancements into the clinical practice will be helpful in better defining the different molecular subsets of melanoma patients and provide new tools to address relevant questions on disease management. Genomic technologies might indeed allow to better predict the biological - and, subsequently, clinical - behavior for each subset of melanoma patients as well as to even identify all molecular changes in tumor cell populations during disease evolution toward a real achievement of a personalized medicine

Computational Integrative Models for Cellular Conversion: Application to Cellular Reprogramming and Disease Modeling

The groundbreaking identification of only four transcription factors that are able to induce pluripotency in any somatic cell upon perturbation stimulated the discovery of copious amounts of instructive factors triggering different cellular conversions. Such conversions are highly significant to regenerative medicine with its ultimate goal of replacing or regenerating damaged and lost cells. Precise directed conversion of damaged cells into healthy cells offers the tantalizing prospect of promoting regeneration in situ. In the advent of high-throughput sequencing technologies, the distinct transcriptional and accessible chromatin landscapes of several cell types have been characterized. This characterization provided clear evidences for the existence of cell type specific gene regulatory networks determined by their distinct epigenetic landscapes that control cellular phenotypes. Further, these networks are known to dynamically change during the ectopic expression of genes initiating cellular conversions and stabilize again to represent the desired phenotype. Over the years, several computational approaches have been developed to leverage the large amounts of high-throughput datasets for a systematic prediction of instructive factors that can potentially induce desired cellular conversions. To date, the most promising approaches rely on the reconstruction of gene regulatory networks for a panel of well-studied cell types relying predominantly on transcriptional data alone. Though useful, these methods are not designed for newly identified cell types as their frameworks are restricted only to the panel of cell types originally incorporated. More importantly, these approaches rely majorly on gene expression data and cannot account for the cell type specific regulations modulated by the interplay of the transcriptional and epigenetic landscape. In this thesis, a computational method for reconstructing cell type specific gene regulatory networks is proposed that aims at addressing the aforementioned limitations of current approaches. This method integrates transcriptomics, chromatin accessibility assays and available prior knowledge about gene regulatory interactions for predicting instructive factors that can potentially induce desired cellular conversions. Its application to the prioritization of drugs for reverting pathologic phenotypes and the identification of instructive factors for inducing the cellular conversion of adipocytes into osteoblasts underlines the potential to assist in the discovery of novel therapeutic interventions

Computational Integrative Models for Cellular Conversion: Application to Cellular Reprogramming and Disease Modeling

The groundbreaking identification of only four transcription factors that are able to induce pluripotency in any somatic cell upon perturbation stimulated the discovery of copious amounts of instructive factors triggering different cellular conversions. Such conversions are highly significant to regenerative medicine with its ultimate goal of replacing or regenerating damaged and lost cells. Precise directed conversion of damaged cells into healthy cells offers the tantalizing prospect of promoting regeneration in situ. In the advent of high-throughput sequencing technologies, the distinct transcriptional and accessible chromatin landscapes of several cell types have been characterized. This characterization provided clear evidences for the existence of cell type specific gene regulatory networks determined by their distinct epigenetic landscapes that control cellular phenotypes. Further, these networks are known to dynamically change during the ectopic expression of genes initiating cellular conversions and stabilize again to represent the desired phenotype. Over the years, several computational approaches have been developed to leverage the large amounts of high-throughput datasets for a systematic prediction of instructive factors that can potentially induce desired cellular conversions. To date, the most promising approaches rely on the reconstruction of gene regulatory networks for a panel of well-studied cell types relying predominantly on transcriptional data alone. Though useful, these methods are not designed for newly identified cell types as their frameworks are restricted only to the panel of cell types originally incorporated. More importantly, these approaches rely majorly on gene expression data and cannot account for the cell type specific regulations modulated by the interplay of the transcriptional and epigenetic landscape. In this thesis, a computational method for reconstructing cell type specific gene regulatory networks is proposed that aims at addressing the aforementioned limitations of current approaches. This method integrates transcriptomics, chromatin accessibility assays and available prior knowledge about gene regulatory interactions for predicting instructive factors that can potentially induce desired cellular conversions. Its application to the prioritization of drugs for reverting pathologic phenotypes and the identification of instructive factors for inducing the cellular conversion of adipocytes into osteoblasts underlines the potential to assist in the discovery of novel therapeutic interventions

A Multi-Objective Genetic Algorithm with Side Effect Machines for Motif Discovery

Understanding the machinery of gene regulation to control gene expression has been one of the main focuses of bioinformaticians for years. We use a multi-objective genetic algorithm to evolve a specialized version of side effect machines for degenerate motif discovery. We compare some suggested objectives for the motifs they find, test different multi-objective scoring schemes and probabilistic models for the background sequence models and report our results on a synthetic dataset and some biological benchmarking suites. We conclude with a comparison of our algorithm with some widely used motif discovery algorithms in the literature and suggest future directions for research in this area

Machine Learning Approaches for Cancer Analysis

In addition, we propose many machine learning models that serve as contributions to solve a biological problem. First, we present Zseq, a linear time method that identifies the most informative genomic sequences and reduces the number of biased sequences, sequence duplications, and ambiguous nucleotides. Zseq finds the complexity of the sequences by counting the number of unique k-mers in each sequence as its corresponding score and also takes into the account other factors, such as ambiguous nucleotides or high GC-content percentage in k-mers. Based on a z-score threshold, Zseq sweeps through the sequences again and filters those with a z-score less than the user-defined threshold. Zseq is able to provide a better mapping rate; it reduces the number of ambiguous bases significantly in comparison with other methods. Evaluation of the filtered reads has been conducted by aligning the reads and assembling the transcripts using the reference genome as well as de novo assembly. The assembled transcripts show a better discriminative ability to separate cancer and normal samples in comparison with another state-of-the-art method. Studying the abundance of select mRNA species throughout prostate cancer progression may provide some insight into the molecular mechanisms that advance the disease. In the second contribution of this dissertation, we reveal that the combination of proper clustering, distance function and Index validation for clusters are suitable in identifying outlier transcripts, which show different trending than the majority of the transcripts, the trending of the transcript is the abundance throughout different stages of prostate cancer. We compare this model with standard hierarchical time-series clustering method based on Euclidean distance. Using time-series profile hierarchical clustering methods, we identified stage-specific mRNA species termed outlier transcripts that exhibit unique trending patterns as compared to most other transcripts during disease progression. This method is able to identify those outliers rather than finding patterns among the trending transcripts compared to the hierarchical clustering method based on Euclidean distance. A wet-lab experiment on a biomarker (CAM2G gene) confirmed the result of the computational model. Genes related to these outlier transcripts were found to be strongly associated with cancer, and in particular, prostate cancer. Further investigation of these outlier transcripts in prostate cancer may identify them as potential stage-specific biomarkers that can predict the progression of the disease. Breast cancer, on the other hand, is a widespread type of cancer in females and accounts for a lot of cancer cases and deaths in the world. Identifying the subtype of breast cancer plays a crucial role in selecting the best treatment. In the third contribution, we propose an optimized hierarchical classification model that is used to predict the breast cancer subtype. Suitable filter feature selection methods and new hybrid feature selection methods are utilized to find discriminative genes. Our proposed model achieves 100% accuracy for predicting the breast cancer subtypes using the same or even fewer genes. Studying breast cancer survivability among different patients who received various treatments may help understand the relationship between the survivability and treatment therapy based on gene expression. In the fourth contribution, we have built a classifier system that predicts whether a given breast cancer patient who underwent some form of treatment, which is either hormone therapy, radiotherapy, or surgery will survive beyond five years after the treatment therapy. Our classifier is a tree-based hierarchical approach that partitions breast cancer patients based on survivability classes; each node in the tree is associated with a treatment therapy and finds a predictive subset of genes that can best predict whether a given patient will survive after that particular treatment. We applied our tree-based method to a gene expression dataset that consists of 347 treated breast cancer patients and identified potential biomarker subsets with prediction accuracies ranging from 80.9% to 100%. We have further investigated the roles of many biomarkers through the literature. Studying gene expression through various time intervals of breast cancer survival may provide insights into the recovery of the patients. Discovery of gene indicators can be a crucial step in predicting survivability and handling of breast cancer patients. In the fifth contribution, we propose a hierarchical clustering method to separate dissimilar groups of genes in time-series data as outliers. These isolated outliers, genes that trend differently from other genes, can serve as potential biomarkers of breast cancer survivability. In the last contribution, we introduce a method that uses machine learning techniques to identify transcripts that correlate with prostate cancer development and progression. We have isolated transcripts that have the potential to serve as prognostic indicators and may have significant value in guiding treatment decisions. Our study also supports PTGFR, NREP, scaRNA22, DOCK9, FLVCR2, IK2F3, USP13, and CLASP1 as potential biomarkers to predict prostate cancer progression, especially between stage II and subsequent stages of the disease