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Acquired resistance to HER2-targeted therapies occurs frequently in HER2+ breast tumors and new strategies for overcoming resistance are needed. Here, we report that resistance to trastuzumab is reversible, as resistant cells regained sensitivity to the drug after being cultured in drug-free media. RNA-sequencing analysis showed that cells resistant to trastuzumab or trastuzumab + pertuzumab in combination increased expression of oxidative phosphorylation pathway genes. Despite minimal changes in mitochondrial respiration, these cells exhibited increased expression of ATP synthase genes and selective dependency on ATP synthase function. Resistant cells were sensitive to inhibition of ATP synthase by oligomycin A, and knockdown of ATP5J or ATP5B, components of ATP synthase complex, rendered resistant cells responsive to a low dose of trastuzumab. Furthermore, combining ATP synthase inhibitor oligomycin A with trastuzumab led to regression of trastuzumab-resistant tumors in vivo. In conclusion, we identify a novel vulnerability of cells with acquired resistance to HER2-targeted antibody therapies and reveal a new therapeutic strategy to overcome resistance.Significance:These findings implicate ATP synthase as a novel potential target for tumors resistant to HER2-targeted therapies.

Supplementary Table 2: BT-TR1 versus BT474 RNA sequencing normalized gene expression Supplementary Table 3: BT-TR2 versus BT474 RNA sequencing normalized gene expression Supplementary Table 4: BT-TPR1 versus BT474 RNA sequencing normalized gene expression Supplementary Table 5: BT-TPR2 versus BT474 RNA sequencing normalized gene expression Supplementary Table 6: BT-TR1 versus BT474 positively enriched hallmark pathways. Supplementary Table 7: BT-TR2 versus BT474 positively enriched hallmark pathways. Supplementary Table 8: BT-TPR1 versus BT474 positively enriched hallmark pathways. Supplementary Table 9: BT-TPR2 versus BT474 positively enriched hallmark pathways. Supplementary Table 10: BT-TR1 versus BT474 negatively enriched hallmark pathways. Supplementary Table 11: BT-TR2 versus BT474 negatively enriched hallmark pathways. Supplementary Table 12: BT-TPR1 versus BT474 negatively enriched hallmark pathways. Supplementary Table 13: BT-TPR2 versus BT474 negatively enriched hallmark pathways.

Supplementary Figure 1: Generation and characterization of resistant cell pools. Supplementary Figure 2: Copy number assessment of receptor tyrosine kinases in resistant cell pools. Supplementary Figure 3: Trastuzumab resistance is reversible. Supplementary Figure 4: Drug resistant cells are more sensitive to OXPHOS inhibitors than parental cells. Supplementary Figure 5: Drug resistant cells have similar mitochondrial DNA copy numbers and ATP levels as the parental cells. Supplementary Figure 6: Glycolysis in parental BT474 versus trastuzumab resistant cells. Supplementary Figure 7: ATP synthase gene expression in BT474, SKBR3 cells and their derived trastuzumab-resistant cells. Supplementary Figure 8: Cumulative survival of breast cancer (all, basal, luminal) patients with patients divided in half by ATP5B (left) or ATP5A1 (right) mRNA level. Supplementary Figure 9: ATP5B protein expression correlation with survival in HER2+, trastuzumab treated patients from HeCOG 10/05. Supplementary Figure 10: ATP5B is required for the maintenance of resistance. Supplementary Figure 11: BT-TR2 tumor morphology and proliferation. Supplementary Table 1: Sequences of primers and plasmid inserts. Supplementary Methods Supplementary References

A síndrome da malformação cavernomatosa múltipla familiar é caracterizada por ser uma herança autossômica dominante de acometimento epidemiológico bimodal. Tem como manifestações clínicas cefaléia, crises convulsivas, hemorragia e déficits neurológicos focais. O diagnóstico é feito por ressonância magnética e o tratamento através de controle dos sintomas, microcirurgia e radiocirurgia. Trata-se de uma patologia rara, mas que pode causar grande impacto na qualidade de vida e mortalidade das famílias acometidas.

Introdução: Em vítimas de trauma abdominal fechado o baço é o órgão mais frequentemente comprometido (40-55%), o mais suscetível a lesões graves e, por ser intensamente vascularizado, pode manter o paciente instável hemodinamicamente, sendo associado com significante morbidade e mortalidade Apresentação do caso: MCRM, sexo feminino, 19 anos de idade, admitida no Hospital Universitário Risoleta Tolentino Neves (UFMG) após queda de escada, queixando dor abdominal. Negou comorbidades e uso de medicamentos. Apresentava-se hemodinamicamente estável, com sons respiratórios diminuídos em bases pulmonares. Discussão: O tratamento quase exclusivo no passado se resumia à laparotomia e esplenectomia. Porém, práticas menos invasivas passaram a ser mais recomendadas com o tempo. Assim, o tratamento padrão-ouro para o trauma esplênico atualmente é o não operatório Conclusão: A principal causa de falha do tratamento é a hemorragia cuja incidência aumenta com o grau da lesão. Nem todos os autores recomendam indiscriminadamente o tratamento conservador para as lesões grau IV (AAST) onde a incidência de hemorragia varia de 33% a 45%. Ficando reservado o tratamento cirúrgico por via laparotômica para as lesões grau IV e V (AAST)

Tumors acquire alterations in oncogenes and tumor suppressor genes in an adaptive walk through the fitness landscape of tumorigenesis. However, the features of this landscape remain poorly understood and cannot be revealed by human cancer genotyping alone. Here, we use a multiplexed, autochthonous mouse platform to model and quantify the initiation and growth of more than one hundred genotypes of lung tumors across four oncogenic contexts: KRAS G12D, KRAS G12C, BRAF V600E, and EGFR L858R. The resulting fitness landscape is rugged (the effect of tumor suppressor inactivation often switches between beneficial and deleterious depending on the oncogenic context), shows no evidence of diminishing-returns epistasis within variants of the same oncogene, and is inconsistent with expectations of a simple linear signaling relationship among these three oncogenes. Our findings suggest that tumor suppressor effects are strongly context-specific, which limits the set of evolutionary paths that can be taken through the fitness landscape.

Este artigo visa apresentar o Grupo de Pesquisa em Empoderamento Feminino (GPEF), uma atividade desenvolvida pelo Programa de Educacao Tutorial do Curso de Engenharia Eletrica da Universidade Federal da Bahia (PET Eletrica UFBA), assim como a metodologia empregada para seu desenvolvimento e os resultados obtidos ate o momento. O grupo de pesquisa tem como objetivo tratar de questoes de genero e, assim, diagnosticar a baixa participacao de mulheres no curso de graduacao em Engenharia Eletrica da UFBA, atraves da analise de dados, e adquirir embasamento teorico para realizacao de atividades extensionistas e acoes em redes sociais. Tendo em vista a baixa participacao feminina nas areas das Ciencias Exatas e Engenharias, com a escrita deste artigo busca-se provocar a criacao de grupos de proposito semelhante a fim de incentivar mulheres nas suas areas de conhecimento.

Cancer progression is a quintessential example of a walk on an adaptive fitness landscape, with tumor growth depending on the cooperation of multiple driver mutations. While sequencing of tens of thousands of clinical samples has revealed a vast set of cancer drivers, far less is known about the interactions of oncogene-tumor suppressor pairs. Due to patient-level selection bias, confounding biological factors and limited sample sizes, a map of these interactions cannot be generated from human data alone and, instead, requires direct perturbational experiments and functional genomics approaches. Here, we model and quantify tumors using an autochthonous mouse platform and Tubaseq, which integrates barcoded lentiviral-sgRNA/Cre vectors and high-throughput barcode sequencing to uncover the number of neoplastic cells in each tumor of each genotype. Across four oncogenic contexts—KRAS G12D, KRAS G12C, BRAF V600E, and EGFR L858R—we analyzed >10,000,000 tumors to estimate the tumorigenesis potential of the oncogene alone as well as the effect of inactivating 28 tumor suppressor genes. We discovered that despite KRAS G12D producing >10x the tumors as G12C, tumor suppressor inactivations provide similar growth effects on both backgrounds. In contrast, although the intrinsic abilities of BRAF V600E and EGFR L858R to drive tumor development fall within the range of the KRAS variants, tumor suppressive effects are categorically different in the context of each oncogene. Many tumor suppressors show clear sign epistasis with the oncogenes, whereby inactivation is advantageous in one context and neutral or deleterious in another. Inactivation of some of the strongest tumor suppressors (e.g., Lkb1, Setd2, and Kmt2d) in KRAS-driven tumors strongly decreases tumor growth in the presence of oncogenic EGFR. While some of these epistatic effects are consistent with a textbook understanding of the RAS pathway, most cannot be predicted based on the linear oncogenic EGFR → KRAS → BRAF pathway model. Analyses of clinical genomics data from AACR Project GENIE confirm that high rates of passenger mutations in KRAS- and BRAF-driven lung tumors, among other factors, prevent the discovery of these interactions from human data alone. However, for EGFR-mutant lung cancers, which are less confounded by high mutational burden, the rates of coincident tumor suppressor mutation are highly correlated with tumor growth effects in our in vivo model. Thus, we find via causal experiments that the landscape of tumor suppression is highly dependent on oncogenic context, with a minority of tumor suppressive effects robust to changes in the oncogene. These findings suggest that the utility of a specific cancer mutation as a prognostic or predictive biomarker of patient outcomes will be dependent on coincident mutations in the tumor and highlight the utility of high-throughput, quantitative autochthonous mouse models in advancing our understanding of cancer biology. Citation Format: Lily M. Blair, Joseph M. Juan, Lafia Sebastian, Vy B. Tran, Wensheng Nie, Gregory D. Wall, Mehmet Gerceker, Ian K. Lai, Edwin A. Apilado, Gabriel Grenot, David Amar, Giorgia Foggetti, Mariana Do Carmo, Zeynep Ugur, Debbie Deng, Alex Chenchik, Maria Paz Zafra, Lukas E. Dow, Katerina Politi, Jonathan J. MacQuitty, Dmitri A. Petrov, Monte M. Winslow, Michael J. Rosen, Ian P. Winters. Oncogenic context shapes the fitness landscape of tumor suppression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1172.

The muscular performance of the lumbar spine is frequently assessed in studies that compare populations with and without low back pain. However, the gold standard equipment used to evaluate the strength of these muscles has a high cost. Therefore, rarely used in the clinical practice of health professionals, being the handheld dynamometer is an accessible alternative for use in clinical practice. Therefore, this study aimed to determine the reliability of the handheld dynamometer in the measurement of maximum voluntary isometric contraction of trunk extensors in healthy adults. Twenty-six healthy adults of both sexes (21.53± 1.88 years old) performed the test and retest with a 1-week interval. The comparison between test-retest demonstrated high and very high intraclass correlation coefficient for peak strength (0.82; 0.60 to 0.92) and mean strength (0.90; 0.76 to 0.95), respectively. Also, the Bland-Altman analyses indicate an error of 5.10 kg for peak strength and 5.7 kg for mean strength. In addition, the minimum detectable change for peak strength was 6.03kg (16.18%) and 5.92 kg for mean strength (20.95%). In conclusion, the test presents high reliability for the extensor muscles of the trunk in healthy adults. Therefore, the use of the handheld dynamometer can be indicated for clinical practice. Keywords: manual dynamometer; assessment; force; back strength; evidence-based practice., O desempenho muscular da coluna lombar é frequentemente avaliado em estudos que comparam populações com e sem lombalgia. Porém, o equipamento padrão-ouro utilizado para avaliar a força desses músculos é de alto custo. Portanto, raramente utilizado na prática clínica de profissionais de saúde, sendo assim o dinamômetro portátil uma alternativa acessível para uso na prática clínica. Portanto, o objetivo deste estudo foi determinar a confiabilidade do dinamômetro portátil na mensuração da contração isométrica voluntária máxima de extensores de tronco em adultos saudáveis. Vinte e seis adultos saudáveis ​​de ambos os sexos (21,53 ± 1,88 anos) realizaram o teste e reteste com intervalo de 1 semana. A comparação entre teste-reteste demonstrou coeficiente de correlação intraclasse alto e muito alto para força de pico (0,82; 0,60 a 0,92) e força média (0,90; 0,76 a 0,95), respectivamente. Além disso, as análises de Bland-Altman indicam um erro de 5,10 kg para força de pico e 5,7 kg para força média. Ademais, a mudança mínima detectável para força de pico foi 6,03kg (16,18%) e 5,92kg para força média (20,95%). Em conclusão, o teste apresenta alta confiabilidade para os músculos extensores do tronco em adultos saudáveis. Portanto, o uso do dinamômetro portátil pode ser indicado para a prática clínica. Palavras-chave: Dinamômetro manual, Avaliação, Força, Força de Extensores do Tronco, Prática Baseada em Evidências.

The ability to understand and predict variable responses to therapeutic agents may improve outcomes in patients with cancer. We hypothesized that the basal gene-transcription state of cancer cell lines, coupled with cell viability profiles of small molecules, might be leveraged to nominate specific mechanisms of intrinsic resistance and to predict drug combinations that overcome resistance. We analyzed 564,424 sensitivity profiles to identify candidate gene-compound pairs, and validated nine such relationships. We determined the mechanism of a novel relationship, in which expression of the serine hydrolase enzymes monoacylglycerol lipase (MGLL) or carboxylesterase 1 (CES1) confers resistance to the histone lysine demethylase inhibitor GSK-J4 by direct enzymatic modification. Insensitive cell lines could be sensitized to GSK-J4 by inhibition or gene knockout. These analytical and mechanistic studies highlight the potential of integrating gene-expression features with small-molecule response to identify patient populations that are likely to benefit from treatment, to nominate rational candidates for combinations and to provide insights into mechanisms of action.

Orientador: Agueda Bernardete Bittencourt Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Educação Resumo: O presente trabalho baseou-se na pesquisa e análise de dados e documentos das escolas de educação infantil da cidade de Campinas, buscando perfazer o histórico de configuração da Rede Escolar de Educação Infantil de Campinas. Para tanto, foi realizado um mapeamento com todas as unidades construídas em suas diversas modalidades, neste trabalho classificadas como Pública Municipal, Pública Conveniada e Pública Privada. Tomando os conceitos de Configuração, Rede e Interdependência, elaborados por Norbert Elias e considerando a relação indissociável entre indivíduo e sociedade, e esta como fruto de uma configuração social proveniente de uma teia de relações de indivíduos interligados e dependentes em densas tramas sociais, construí a interpretação da Rede Pública de Educação Infantil de Campinas. Dois eixos nortearam minhas interpretações: as relações entre educação infantil e assistência social; educação infantil e a religião. Foram tomados como fonte de pesquisa os relatórios estatísticos oficiais publicados pela Prefeitura Municipal de Campinas, sites da prefeitura e, projetos políticos pedagógicos das unidades escolares, além das Diretrizes Curriculares Municipais da Educação Básica para o Ensino Fundamental ¿ Anos Iniciais e para a Educação Infantil, publicadas em 2012 e 2013 respectivamente Abstract: The present work was based on the research and analysis of data and documents of early childhood schools in the city of Campinas, seeking to make up the history of configuration of the Early Childhood Education Network of Campinas. For this purpose, a mapping was carried out with all the units built in their different modalities, in this work classified as Public Municipal, Public Partnership and Public Private. Taking the concepts of Configuration, Network and Interdependence, developed by Norbert Elias and considering the inseparable relationship between individual and society, and this as the result of a social configuration arising from a web of relationships of interconnected and dependent individuals in dense social networks, I built the interpretation of the Public Child Education Network of Campinas. Two axes guided my interpretations: the relationship between early childhood education and social assistance; child education and religion. The official statistical reports published by the City of Campinas, websites of the city hall and pedagogical political projects of the school units were taken as a source of research, in addition to the Municipal Curriculum Guidelines for Basic Education for Elementary Schools - Early Years and for Early Childhood Education, published in 2012 and 2013 respectively Mestrado Educação Mestra em Educação

Acquired resistance to HER2-targeted therapies occurs frequently in HER2+ breast tumors and new strategies for overcoming resistance are needed. Here, we report that resistance to trastuzumab is reversible, as resistant cells regained sensitivity to the drug after being cultured in drug-free media. RNA-sequencing analysis showed that cells resistant to trastuzumab or trastuzumab + pertuzumab in combination increased expression of oxidative phosphorylation pathway genes. Despite minimal changes in mitochondrial respiration, these cells exhibited increased expression of ATP synthase genes and selective dependency on ATP synthase function. Resistant cells were sensitive to inhibition of ATP synthase by oligomycin A, and knockdown of ATP5J or ATP5B, components of ATP synthase complex, rendered resistant cells responsive to a low dose of trastuzumab. Furthermore, combining ATP synthase inhibitor oligomycin A with trastuzumab led to regression of trastuzumab-resistant tumors in vivo. In conclusion, we identify a novel vulnerability of cells with acquired resistance to HER2-targeted antibody therapies and reveal a new therapeutic strategy to overcome resistance. Significance: These findings implicate ATP synthase as a novel potential target for tumors resistant to HER2-targeted therapies.

Drug resistant mutations that arise in therapeutic targets often limit clinical responses. However, the discovery of such mutations has historically been performed one gene or mutation at a time, often over decades of experimental and clinical testing, limiting our understanding of conserved mechanisms of drug resistance. We hypothesized that deep mutational scanning of canonical kinases may expedite this process and identify novel conserved elements that cause drug resistance when mutated (similar to the well-studied “Gatekeeper” residue). To test this, we generated cDNA-expression libraries containing all possible amino acid substitutions in CDK6, CDK4, ERK2, and EGFR. We screened each library against clinically utilized, ATP-competitive small molecule inhibitors. We then mapped the phenotypic data for over 40,000 missense mutations onto the aligned crystal structures of each protein and searched for shared structural attributes associated with drug resistance. This analysis revealed 4 equivalent amino acid sites whose mutation conferred drug resistance to ATP-competitive inhibitors in all of our screens: the Gatekeeper residue, as well as three uncharacterized residues. One of these sites, which we have termed the “Keymaster”, was additionally found to cause resistance in published data sets of sub-saturation BRAF, HER2, BCR-ABL, and MEK1 mutagenesis screens against their respective inhibitors. We confirmed that drug resistant phenotypes are caused by these alterations utilizing growth assays and protein target phosphorylation detection assays. Mechanistically, we show preliminary evidence that Keymaster-mutant proteins are competent for drug binding, but may display elevated basal activity. Consistent with our findings, we additionally identified mutations at Keymaster residues in reported patient tumors in a number of oncogene kinases, suggesting that Keymaster mutations could be drivers of tumorigenesis, as well as drug resistance. These efforts may prove useful for characterizing somatic kinase mutations of unknown function, designing next-generation therapeutics and deepening our understanding of kinase regulation. Citation Format: Nicole S. Persky, Desiree Hernandez, Jonathon Cordova, Amanda Walker, Lisa Brenan, Federica Piccioni, Sasha Pantel, Yenarae Lee, Amy Goodale, Xiaoping Yang, Yoichiro Mitsuishi, Mariana Do Carmo, Cong Zhu, Aleksandr Andreev, David E. Root, Cory M. Johannessen. Massively parallel identification of conserved drug resistant mutations in kinases [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1815.

Dissertação (mestrado) - Pontifícia Universidade Católica de Minas Gerais, Programa de Pós-Graduação em Psicologia. Bibliografia: f. 104-110 Resumo: O objeto de estudo da presente pesquisa foram as tensões que os psicólogos vivem atualmente no Centro de Referência Especializado de Assistência Social (CREAS), unidade da proteção social especial de média complexidade do Sistema Único de Assistência Social (SUAS), bem como as contradições entre o cotidiano de trabalho e o que está instituído no SUAS, examinando quais forças instituintes estão presentes nessas contradições. O objetivo da pesquisa foi investigar e conhecer o trabalho do psicólogo no campo da assistência social, considerando os conflitos encontrados por este profissional em relação à demanda real de trabalho e o que está instituído por lei e pela política do SUAS, através da Política Nacional de Assistência Social (PNAS) e sua legislação. Na pesquisa de campo, foram realizadas entrevistas coletivas semiestruturadas em três das nove regionais da Secretaria Municipal Adjunta de Assistência Social da Prefeitura de Belo Horizonte, além da utilização do diário de campo e da implicação da pesquisadora. Para uma melhor compreensão do contexto pesquisado, fez-se necessário descrever os princípios norteadores da política pública de Assistência Social com enfoque na especificidade do CREAS; examinar a intervenção do psicólogo no que se refere ao campo de forças do instituído e do instituinte e sua relação dialética, a partir da Análise Institucional de René Lourau; e analisar os possíveis desdobramentos resultantes dessas intervenções em relação à proposta de atuação no CREAS. Através da Análise de Conteúdo proposta por Bardin, foram escolhidas as seguintes categorias, também chamadas de analisadores: entrada e saída do SUAS, dinâmica do trabalho e formação em Psicologia. Concluiu-se que este ainda é um campo em construção, com um forte instituído cristalizado historicamente, tanto pelo modo de funcionamento da assistência social em nosso país quanto pela própria formação recebida nos cursos de graduação, que já começam a pensar na sustentação de novas práticas que se abram para as forças instituintes. Abstract: The object of study of this research were the tensions that psychologists currently live in Specialized Reference Center for Social Assistance (CREAS), a unit of special social protection of average complexity of the Unified Social Assistance System (SUAS), as well as the contradictions between the daily work and what is established in the SUAS, examining which instituting forces are present in these contradictions. The objective of the research was to investigate and know the psychologist's work in the field of social assistance, considering the conflicts encountered by this professional compared to the real demand for labor and what is established by law and by the SUAS policy, through the National Social Assistance Policy (PNAS) and its legislation. In field research, semi-structured group interviews were carried out in three of the nine regions of the Municipal Assistant Secretary for Social Assistance of the Municipality of Belo Horizonte, in addition to the use of the field diary and the implication of the researcher. For a better understanding of the researched context, it was necessary to describe the guiding principles of public policy Social Assistance focusing on the specificity of CREAS; to examine the intervention of the psychologist regarding the force field of instituted and instituting and its dialectical relationship, according to René Lourau Institutional Analysis; and to analyze the possible consequences resulting from these interventions in relation to the actions proposed in CREAS. Through Content Analysis proposed by Bardin, the following categories also called analyzers were chosen: admission and leaving of the SUAS, dynamics of work and training in Psychology. It was concluded that this field is still under construction, with a strong instituted historically crystallized, both by the social assistance operating mode in our country and by the training received in undergraduate courses, which are beginning to think about the support of new practices that are open to the instituting forces. O CD-ROM que acompanha a obra encontra-se no setor de Coleções Especiais/Audiovisual da Biblioteca

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