Objetivo: Determinar el perfil epidemiológico del cáncer de mama HER2- positivo en pacientes atendidos en el Hospital Nacional Hipólito Unanue entre Junio del 2012 a Junio del 2015. Material: revisión retrospectiva de fichas de notificación, (Directiva Sanitaria de Vigilancia Epidemiológica del Cáncer. Registros Hospitalarios) de 34 pacientes con cáncer de mama HER 2 – positivo entre Junio del 2012 a Junio del 2015, en el Hospital Nacional Hipólito Unanue, en la Unidad de cáncer de mama y patología mamaria. Resultados: la edad media fue de 48 años (11,76%), el tamaño tumoral más frecuente fue de 2 a 5 cm (47,06%), el grado histológico más frecuente corresponde a pobremente diferenciado (64,71%), el mayor porcentaje (61,76%) no tuvo recurrencia local, el mayor porcentaje (94,12%) no tuvo metástasis, el tratamiento más frecuente fue el de terapia doble (50,00%), que incluye mastectomía radical y quimioterapia. Conclusiones: La clasificación del cáncer de mama basada en parámetros inmunohistoquímicos (IHQ) permite una mejor definición del pronóstico.

Introduction : De nombreuses thérapeutiques ciblant la protéine HER2 ou des protéines clés de la voie HER2 ont transformé le pronostic des cancers du sein HER2 positif. Les anomalies moléculaires tumorales, la richesse des thérapeutiques actuelles et leur coût nécessitent de rationnaliser la décision thérapeutique par l’utilisation de biomarqueurs. La protéomique ciblée, capable de détecter et mesurer un panel de protéines au sein d’échantillons complexes, est l’une des approches les plus performantes pour quantifier un jeu de biomarqueurs spécifiques simultanément dans un tissu tumoral. Objectif : développer une approche de protéomique quantitative ciblée de type PRM (Parallel Reaction Monitoring) pour évaluer les protéines clés de la voie HER2 dans différents échantillons tumoraux. Résultats : 1/ Sélection des peptides protéotypiques de nos protéines d’intérêt (EGFR, HER2 et phospho-HER2, HER3, PTEN) et développement des méthodes d’analyse. 2/ Détection et quantification de ces peptides a/ dans 17 lignées cellulaires mammaires, en conditions standard et après exposition au trastuzumab ou lapatinib. b/ dans des xénogreffes dérivées de cancer du sein humain. 4/ Corrélation des résultats a/ aux « gold » standards actuels : western blot et immuno-cyto/histo-chimie, b/ aux données issues d’analyses transcriptomiques validées. Conclusion : cette approche pourrait permettre dans le futur une vision globale de l’expression et l’activation des protéines de la voie HER2 et progresser vers une médecine « personnalisée ». Elle pourrait être améliorée par l’utilisation de spectromètres de masse plus performants et le recours à la microdissection laser sur tissu conservé en FFPE. / Therapeutics targeting HER2 protein or its pathway considerably improved the prognosis of HER2-positive BC. The actual approaches to evaluate HER2 expression, immunohistochemistry (IHC) and FISH (fluorescent in situ hybridization), are labor-intensive and low-throughput, and present discrepancies between them. Therefore, there is a real need to develop other strategies to rationalize the use of targeted therapeutics. Parallel Reaction Monitoring (PRM) is a mass spectrometry-based approach for targeted proteomics able to detect and quantify numerous proteins with high-throughput allowing to follow mutated or activated status of targeted proteins. PRM could be a way to rationalize the use of targeted therapeutics to go through a more « personalized » medicine. The objective was to detect and quantify proteins implicated in HER2 pathway (EGFR, HER2, HER3, PTEN), phosphorylated peptides of HER2 and their response under treatment. We first selected proteotypic peptides of each protein and protein assays were generated. We detected and quantified proteotypic peptides of proteins of interest 1/on 17 breast cell lines on control condition and under treatment (trastuzumab or lapatinib). 2/ on more complex samples including patients-derived xenografts and human breast cancers. We correlated our data to gold standards western blot and IHC and to transcriptomic signatures previously validated. In the future, this approach could envision a picture of the expression and activation of proteins implicated in HER2-pathway. However, our strategy could be improved by more efficient mass spectrometers and the use of formalin-fixed paraffin-embedded samples to be used in clinical practice.

A Thesis submitted to The University of Arizona College of Medicine - Phoenix in partial fulfillment of the requirements for the Degree of Doctor of Medicine. / Recent advances in cancer immunology, highlighted by immune checkpoint inhibitors, have demonstrated that immunotherapy is a viable option in the oncologist’s armamentarium. Despite these advances, many patients are nonresponders. Preliminary studies have suggested that non-responders lack a de-novo anti-tumor antigen immune response that can be unmasked by checkpoint blockade; thus, strategies to induce anti-tumor immune responses are needed. We hypothesized that many tumor associated antigens (Ag) are readily susceptible to immune attack, but only in the context of identifying the tumor antigen epitopes that can reliably initiate an immune response, regardless of individual patient human leukocyte antigen (HLA) haplotype restrictions. We further hypothesized that epitope prediction strategies which seek to identify pan- or highly promiscuous-HLA binding epitopes would reduce the number of potential candidates and be more likely to accurately identify high-priority tumor Ag epitopes. Utilizing known HLA-serotype frequencies and setting a threshold of ninety percent of population coverage, regardless of race or ethnicity, twenty-nine different HLA-DRB1 haplotypes were chosen for antigen prediction utilizing the open source epitope prediction algorithm netMHCIIpan. Predictions were also performed for HLA-A serotypes utilizing the open source algorithm netMHCpan. Predicted epitopes were synthesized in the form of synthetic long peptides and tested in immune system sensitization assays involving unfractionated peripheral blood mononuclear cells (PBMC). Briefly, PBMC were subjected to a two-step culture, first synchronizing their exposure to the long peptides with aggressive surrogate activation of innate immunity, followed by IL-7-modulated T-cell hyperexpansion. Predictions resulted in identification of highly promiscuous-HLA binding epitopes. Unexpectedly, these epitopes clustered together forming high priority regions: unique “hot spots” with high densities of promiscuous HLA-binding epitopes from the widely expressed oncoproteins MUC1, HER2/neu and CMV-pp65 (p<0.0001, for predicted HLA-DRB1 binding affinities, compared to non-hot spot regions). Added synthetic long peptides (>20aa) derived from “hot spot” regions of MUC1, HER2/neu, and CMVpp65 reliably produced selective and sustained expansion of both CD4+ and CD8+ peptide-specific, interferon-γ (IFNγ)-producing Tcells when synchronized with step 2 exposure to exogenous IL-7 (p<0.0001 and p=0.0048, for CD4+ and CD8+ Ag-specific T-cells, respectively, compared to T-cells directed against peptides from non-hot spot regions). “Hot spot” peptide Ag-specific T-cells preferentially recognized endogenous tumor derived MUC1, either in MUC1 expressing tumor cell killing assays (p=0.038, compared to non-peptide Ag-specific T-cells) or as MUC1 tumor lysate when pulsed onto restimulatory PBMC (p=0.022 and 0.025, for CD4+ and CD8+ T-cells, respectively, compared to T-cells directed against peptides from non-hot spot regions). This mechanistically rational antigen selection sequence, effective even for unvaccinated donors, regardless of HLA-haplotype, enables rapid identification of tumor protein regions relevant for cancer immunology, including adoptive immunotherapy, vaccines, and even identification of tumor neo-antigens unique to each patient.

MSc (Med), Faculty of Health Sciences, University of the Witwatersrand / The HER2 gene is amplified in 20-30% of breast cancers, a common cancer amongst South African women. HER2 amplification is associated with a poor prognosis and predicts response to treatments such as Herceptin. The gold standard for HER2 testing is Fluorescent in situ Hybridisation (FISH) with dual colour probes for the HER2 gene and chromosome 17 centromere (CEP17) internal control. According to international guidelines, a HER2/CEP17 ratio >2.2 is considered positive. The HER2 FISH test is complicated by the emergence of ambiguous cases with increased CEP17 signals that cannot be accounted for by chromosome 17 polysomy (> 6 copies of CEP17) and that may hide true HER2 gene amplification. The aims of this study were to characterise the HER2 amplicon, in particular the copy number of genes in the vicinity of the HER2 gene, and to design an alternative control probe that could clarify the HER2 gene status in ambiguous cases. In addition, results on 1558 breast cancer specimens sent for routine testing were analysed to determine the trends of HER2 amplification amongst South African women. The rate of HER2 gene amplification was significantly higher (p < 0.05) in African patients (52%) than in Caucasian patients (43%). In Caucasian women, the rate of HER2 amplification in the younger group (68%) was significantly higher (p < 0.05) than in the general Caucasian group (43%), while the same was not seen in the African cohort. Nineteen ambiguous cases with more than 9 copies of CEP17 were further investigated. FISH assays with four different probe kits (PathVysion HER-2: Poseidon Repeat free TOP2A, HER2, CEP17: and Vysis PML-RARA respectively) were performed to determine the copy number of the HER2, TOP2A, RARA genes and CEP17. An in-house dual colour probe kit was designed using the ACTG1 gene as a control for HER2. Of the 19 ambiguous cases, 16 had centromeric amplification, showing that CEP17 is no longer an adequate internal control in FISH HER2 testing. The TOP2A gene was only amplified in HER2 positive cases and the RARA gene was only amplified when the TOP2A gene was also amplified. FISH with ACTG1 as v a control clearly revealed HER2 amplification in ambiguous cases on image analysis and gave HER2/ACTG1 ratios significantly higher than HER2/CEP17 ratios. However, screening of an additional 40 unambiguous cases showed an increased copy number, although limited ( 8), of the ACTG1 gene in four patients; this warrants further testing to assess the value of this gene as a control. Interestingly, a trend was observed for ACTG1 increased copy number in HER2 negative cases, this may point to the presence of a driver gene whose amplification tends to be mutually exclusive from HER2 amplification.

Over the past two decades, the mammalian Ste20-like kinase (SLK) has been characterized for its role in regulating cellular migration, proliferation and apoptosis in fibroblasts and myoblasts. In mammary epithelial cells, SLK has been shown to be required for efficient epithelial-to-mesenchymal transition and to be activated downstream of the HER2/Neu-oncogene to control chemotactic cellular migration. Here, we assessed the role of SLK in HER2/Neu-induced mammary tumorigenesis in vivo. As SLK is activated downstream of HER2/Neu, we hypothesized that the loss of SLK would significantly delay tumor progression in a mouse model of HER2-positive breast cancer. As we have shown that global attenuation of SLK kinase activity results in embryonic lethality, a conditional SLK knockout mouse model was generated. To study the role of SLK in HER2-positive breast cancer, we crossed these conditional SLK knockout mice with mice expressing HER2/Neu linked to Cre recombinase in the mammary luminal epithelium. Unexpectedly, we have demonstrated that conditional deletion of SLK significantly accelerates Neu-induced mammary tumor onset and decreases overall survival. SLK deletion results in the induction of Sox10 which drives mammary stem/progenitor activity and cooperates with HER2/Neu to drive tumor growth. Using the Cancer Genome Atlas, we have supported previous findings and validated Sox10 as a potential biomarker of the Triple-negative Breast Cancer subtype. Furthermore, we have uncovered that SLK deletion results in enhanced activation of both PDK1 and AKT. We provide evidence that Sox10 induction requires signaling through a novel AKT/Sox9-dependent pathway following SLK deletion. Taken together, our data suggests that SLK may be required to maintain cells in a fully differentiated state and that loss of SLK in HER2/Neu-induced breast cancer drives a more basal/stem-like phenotype through the induction of Sox10.

<p>During oncogenesis, cancer cells go through metabolic reprogramming to maintain their high growth rates and adapt to changes in the microenvironment and the lack of essential nutrients. Several types of cancer are dependent on de novo fatty acid synthesis to sustain their growth rates by providing precursors to construct membranes and produce vital signaling lipids. Fatty acid synthase (FASN) catalyze the terminal step of de novo fatty acid synthesis and it is highly expressed in many types of cancers where it’s up-regulation is correlated with cancer aggressiveness and low therapeutic outcome. Many FASN inhibitors were developed and showed potent anticancer activity however, only one inhibitor advanced to early stage clinical trials with some dose limiting toxicities. Using a modified fluorescence-linked enzyme chemoproteomic strategy (FLECS) screen, we identified HS-106, a thiophenopyrimiden FASN inhibitor that has anti-neoplastic activity against breast cancer in vitro and in vivo. HS-106 was able to inhibit both; purified human FASN activity and cellular fatty acid synthesis activity as evaluated by radioactive tracers incorporation into lipids experiments. In proliferation and apoptosis assays, HS-106 was able to block proliferation and induce apoptosis in several breast cancer cell lines. Several rescue experiment and global lipidome analysis were performed to probe the mechanism by which HS-106 induces apoptosis. HS-106 was found to induce several changes in lipids metabolism: (i) inhibit fatty acids synthesis. (ii) Inhibit fatty acids oxidation as indicated by the ability of inhibiting Malonyl CoA accumulation to block HS-106 induced apoptosis and the increase in the abundance of ceramides. (iii) Increase fatty acids uptake and neutral lipids formation as confirmed 14C Palmitate uptake assay and neutral lipids staining. (iv)Inhibit the formation of phospholipids by inhibiting de novo fatty acid synthesis and diverting exogenous fatty acids to neutral lipids. All of these events would lead to disruption in membranes structure and function. HS-106 was also tested in Lapatinib resistant cell lines and it was able to induce apoptosis and synergizes Lapatinib activity in these cell lines. This may be due the disruption of lipid rafts based on the observation that HS-106 reduces the expression of both HER2 and HER3. HS-106 was found to be well tolerated and bioavailable in mice with high elimination rate. HS-106 efficacy was tested in MMTV neu mouse model. Although did not significantly reduced tumor size (alone), HS-106 was able to double the median survival of the mice and showed potent antitumor activity when combined with Carboplatin. Similar results were obtained when same combinations and dosing schedule was used in C3Tag mouse model except for the inability of HS-106 affect mice survival.</p><p>From the above, HS-106 represent a novel FASN inhibitor that has anticancer activity both in vivo and in vitro. Being a chemically tractable molecule, the synthetic route to HS-106 is readily adaptable for the preparation of analogs that are similar in structure, suggesting that, the pharmacological properties of HS-106 can be improved.</p> / Dissertation

Gastroesophageal cancer is the sixth leading cause of cancer-related death worldwide. The 2 most common histologies are squamous cell carcinoma and adenocarcinoma, which has seen an increase in incidence correlating with an increase in obesity in developed countries. Gastroesophageal adenocarcinoma has a preponderance to metastasize early, making it a highly lethal cancer with a low 5-year survival rate of similar to 15-25%. Therefore, for the majority of patients, treatment focuses on palliation and prolongation of survival. Combination chemotherapy regimens, mostly platinum-based, have only modestly prolonged survival in patients with stage IV disease. Recently, it was discovered that the activation of the HER2 receptor plays an important role in a minority of adenocarcinomas of the distal esophagus and stomach. This introduced the treatment option of trastuzumab (Herceptin), a monoclonal antibody directed at the HER2 receptor, which has demonstrated improvement in overall and progression-free survival as noted in the ToGA trial. Currently, the role of Herceptin therapy beyond first-line therapy and outside of combination regimens is not well established. In this case report we review 2 cases of patients with gastroesophageal cancer, with HER2 overexpression, who achieved a robust response to trastuzumab in combination with chemotherapy and were able to maintain a durable response with maintenance trastuzumab monotherapy. (c) 2017 The Author(s) Published by S. Karger AG, Basel.

Breast cancer is the most common tumor disease among women in Sweden. About 7000persons, having a median age of 65, are diagnosed each year with this disease. Withmammography screening, breast cancer can be detected in an early stage which improves theoverall survival (OS). 20-30 % of the breast cancer tumors are overexpressing humanepidermal growth factor receptor 2 (HER2), which is a protein that stimulates cell proliferation.Trastuzumab (Herceptin®) is a humanised monoclonal antibody that targets the HER2-proteinand prevent the signals for cell proliferation.Trastuzumab has earlier been used for treatment of metastatic breast cancer. In the year of 2007trastuzumab was approved for adjuvant treatment of patients who has been medicated withsurgery and/or radiation.The aim of this study was to investigate the effects of adjuvant treatment with trastuzumab inHER2-positive breast cancer patients. The method was a literature study based on scientificarticles identified from the database PubMed.The articles that were choosen were two meta-analysis and three cohort studies. The benefitsand effects of trastuzumab administration on patients with HER2-positive breast cancer weredescribed in these articles. Primary endpoint was disease-free survival (DFS). All articlesshowed that the DFS increased in breast cancer patients treated with adjuvant trastuzumab but alonger follow-up of four years, showed a reduction of both DFS and OS.The conclusion of this study was that trastuzumab as adjuvant treatment is favorable and shouldbe considered as treatment of breast cancer with HER2 overexpression that has been analyzedby immunohistochemistry (IHC) or fluorescence in situ hybridization (FISH).

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20799号 / 医博第4299号 / 新制||医||1025(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 竹内 理, 教授 武藤 学, 教授 河本 宏 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Breast cancer is one of the deadliest forms of cancers in the world. About 15–30% of all breast cancers are HER2-positive, which involves overexpression of HER2-receptors on the surface of tumor cells. HER2-positive breast cancer is viewed as an aggressive form of cancer because the overexpression of the HER2-receptor causes dimerization with other receptors of the same family; together they bind to ligands and become activated. The activation causes fast, uncontrolled cell proliferation that often results in the formation of a tumor. Cancer can be divided into different stages, at stage 4 the cancer cells from the original tumor breaks away, follows the bloodstream, and forms metastases in other places of the body. When cancer evolves and becomes metastatic the prognosis drastically worsens, and the treatment options are limited. Patients often need several lines of cancer treatment. The first line of treatment is usually trastuzumab in combination with pertuzumab and a taxane, second line is usually trastuzumab emtansine. There is no conclusive third line treatment for HER2-positive metastatic breast cancer (MBC). Due to the development of new anti-HER2 treatments over the last two decades, less patients are dying from breast cancer, however most patients diagnosed with HER2-positive MBC are estimated to face an early death.  The objective of this study was to analyze the pharmaceutical advances that has improved the prognosis for patients diagnosed with HER2-positive MBC. The material, on which this study was based on, was obtained from the Pubmed database via the Linnaeus University Library. Five articles were chosen based on criteria relevant to the topic. The articles were published between 2001 and 2021; all of them were randomized controlled trials (RCT).  The subject of the articles was to compare the efficacy and safety of different forms of anti-HER2 treatments, using patient populations diagnosed with HER2-positive MBC. Mainly the patients' disease progression, treatment response and survival time was analyzed. Pharmaceutical safety was assessed by the rate of adverse events. A total of 2513 patients participated in the studies. Among all the different treatment options that were analyzed in the articles, one treatment combination yielded some of the best results. Pyrotinib in combination with capecitabine increased the disease progression-free time, had the highest proportion of patients who responded to treatment as well as the highest proportion of patients with size-reducing lesions for the longest time. However, the patient group receiving pyrotinib also had the highest incidence of serious adverse events and had the largest percentage of patients who chose to discontinue the study due to adverse events.  Analysis of the five articles concludes that the prognosis for patients diagnosed with HER2-positive MBC has been improved by pharmaceutical advances regarding tyrosine kinase inhibitors, pan-HER inhibitors, combination therapy with monoclonal antibodies (single, mixed, modified or conjugated with other drugs) and chemotherapy with different mechanisms of action. The results from the studies indicated that treatment with a single anti-HER2 drug had the lowest effectiveness, and that some drug combinations had better synergistic effects than others, reflecting on patient survival data. Despite the pharmaceutical advances of the past two decades, the prognosis for HER2-positive MBC can still be considered bleak due to its high death rate. Resistance to anti-HER2 drugs is an ongoing concern that requires more research and development of new treatments.