Human microbiota and breast cancer : is there any relevant link? A literature review and new horizons toward personalised medicine

Copyright © 2021 Alpuim Costa, Nobre, Batista, Ribeiro, Calle, Cortes, Marhold, Negreiros, Borralho, Brito, Cortes, Braga and Costa. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.Breast cancer (BC) is the most common malignancy and the second cause of cancer-specific death in women from high-income countries. Recently, gut microbiota dysbiosis emerged as a key player that may directly and/or indirectly influence development, treatment, and prognosis of BC through diverse biological processes: host cell proliferation and death, immune system function, chronic inflammation, oncogenic signalling, hormonal and detoxification pathways. Gut colonisation occurs during the prenatal period and is later diversified over distinct phases throughout life. In newly diagnosed postmenopausal BC patients, an altered faecal microbiota composition has been observed compared with healthy controls. Particularly, β-glucuronidase bacteria seem to modulate the enterohepatic circulation of oestrogens and their resorption, increasing the risk of hormone-dependent BC. Moreover, active phytoestrogens, short-chain fatty acids, lithocholic acid, and cadaverine have been identified as bacterial metabolites influencing the risk and prognosis of BC. As in gut, links are also being made with local microbiota of tumoural and healthy breast tissues. In breast microbiota, different microbial signatures have been reported, with distinct patterns per stage and biological subtype. Total bacterial DNA load was lower in tumour tissue and advanced-stage BC when compared with healthy tissue and early stage BC, respectively. Hypothetically, these findings reflect local dysbiosis, potentially creating an environment that favours breast tumour carcinogenesis (oncogenic trigger), or the natural selection of microorganisms adapted to a specific microenvironment. In this review, we discuss the origin, composition, and dynamic evolution of human microbiota, the links between gut/breast microbiota and BC, and explore the potential implications of metabolomics and pharmacomicrobiomics that might impact BC development and treatment choices toward a more personalised medicine. Finally, we put in perspective the potential limitations and biases regarding the current microbiota research and provide new horizons for stronger accurate translational and clinical studies that are needed to better elucidate the complex network of interactions between host, microorganisms, and drugs in the field of BC.info:eu-repo/semantics/publishedVersio

DNA methylation transcriptionally regulates the putative tumor cell growth suppressor ZNF677 in non-small cell lung cancers

In our study, we investigated the role of ZNF677 in non-small cell lung cancers (NSCLC). By comparing ZNF677 expression in primary tumor (TU) and in the majority of cases also of corresponding non-malignant lung tissue (NL) samples from > 1,000 NSCLC patients, we found tumor-specific downregulation of ZNF677 expression (adjusted p-values < 0.001). We identified methylation as main mechanism for ZNF677 downregulation in NSCLC cells and we observed tumor-specific ZNF677 methylation in NSCLC patients (p < 0.0001). In the majority of TUs, ZNF677 methylation was associated with loss of ZNF677 expression. Moreover, ZNF677 overexpression in NSCLC cells was associated with reduced cell proliferation and cell migration. ZNF677 was identified to regulate expression of many genes mainly involved in growth hormone regulation and interferon signalling. Finally, patients with ZNF677 methylated TUs had a shorter overall survival compared to patients with ZNF677 not methylated TUs (p = 0.013). Overall, our results demonstrate that ZNF677 is trancriptionally regulated by methylation in NSCLCs, suggest that ZNF677 has tumor cell growth suppressing properties in NSCLCs and that ZNF677 methylation might serve as prognostic parameter in these patients

Cancer stem cell maintenance in prostate cancer

Hintergrund: Das Prostatakarzinom ist die häufigste Krebsart und die zweithäufigste krebsassoziierte Todesursache bei Männern. Die Erkrankung wurde aufgrund ihrer Abhängigkeit von mitogenen Signalen des Androgenrezeptors als hormonabhängig beschrieben. Der Großteil der Patienten, die am Prostatakarzinom erkranken, spricht daher auf antihormonale Therapiemodalitäten wie chemische Kastration an. Ein signifikanter Anteil der Patienten entwickelt jedoch kastrationsresistente Tumore, welche, trotz großer rezenter Fortschritte in der Behandlung dieser Erkrankung, mit einem mittleren Gesamtüberleben von rund 18 Monaten ab Diagnose mit unheilbarem und letztlich letalem Krankheitsverlauf verbunden sind. In den letzten Jahren konnten spezialisierte Zellklone innerhalb der intratumoralen Heterogenität von Prostatakarzinomen identifiziert werden, welche Fähigkeiten von Stammzellen wie hohes Proliferationspotential und Multipotenz aufweisen und als Krebsstammzellen bezeichnet werden. Mithilfe eines transgenen Mausmodelles und humanen Zelllinien des Prostatakarzinoms konnten wir im Zuge dieses Dissertationsprojektes eine differentielle Antwort von Prostatakarzinomstammzellen auf Hypoxie nachweisen, welche ihr Überleben zulasten von Proliferation per Deregulation der AKT/mTOR-Signaltransduktionsachse förderten. Methodologie: Das transgene, über t-Antigene des Simian Virus 40 getriebene, Modell der Mausprostata (TRAMP) wurde im Zuge dieser Studie als Quelle für primäre Prostatatumore verwendet. Zucht und Haltung erfolgten im biomedizinischen Zentrum unserer Institution. Einzelzellsuspensionen wurden per enzymatischem Gewebeverdau hergestellt und aus ihnen per Durchflusszytometrie stammzellartige Krebszellen isoliert. Intrazellulärer Proteinnachweis erfolgte ebenfalls mithilfe dieser Methode. Weiters wurden etablierte Prostatakarzinomzelllinien murinen und humanen Ursprungs unter standardisierten und sterilen Zellkulturbedingungen gehalten. Teile der Zellexperimente wurden in hypoxischem Milieu und unter pharamakologischer Beeinflussung von mTOR und HIF1[alpha]-Signalachsen durchgeführt. Weitere Experimentalistik umfasste klassische nukleomische und proteomische Nachweismethoden wie etwa PCR-Techniken und Western Blot. Zellviabilität wurde mittels MTT-Assays analysiert. Spheroidbildung wurde in aus Matrigel bestehenden Matrices durchgeführt und mittels Mikroskop manuell ausgewertet. Apoptose- und Viabilitätsuntersuchungen umfassten zudem AnnexinV-Assays, welche durchflusszytometrisch analysiert wurden. Xenografts wurden in NOD SCID Gamma-Mäusen durchgeführt. Alle Tierexperimente wurden unter Einhaltung tierethischer Richtlinien unserer Institution und nach positivem Votum der Tierethikkommission durchgeführt. Ergebnisse: Prostatakrebsstammzellen aus dem basalen Kompartment des Prostataepithels von primären TRAMP-Tumoren als auch aus humanen und murinen Zelllinien wurden auf Hochregulierung von klassischen krebsstimulierenden molekularen Signalwegen untersucht. Der Transskriptionsfaktor HIF1[alpha] zeigte sich in Krebsstammzellen - verglichen mit nicht-stammzellartigen Krebszellen - überexprimiert und zudem verstärkt stabilisiert. Ferner waren auch bekannte HIF-Zielgene in hypoxischen CSCs überexprimiert und eine durch HIF1[alpha] verursachte Überaktivierung von AKT konnte durch einen molekularen Mechanismus mittels mTOR/S6K/IRS-1-Feedbackloop nachgewiesen werden. Diese Deregulation des AKT/mTOR-Signalweges korrelierte zudem mit primärer Resistenz der stammzellartigen Krebszellen gegenüber pharmakologischer mTOR-Inhibition. Konklusion: Unsere Ergebnisse decken einen Krebsstammzellen inhärenten molekularen Mechanismus auf, welcher ihnen unter hypoxischen Bedingungen über Deregulation von AKT und mTOR einen Überlebensvorteil gegenüber umgebenden Zellen zulasten von Wachstum ermöglicht. Über diesen molekularen Schalter, so unsere abschließende Hypothese, besitzen Prostatakrebsstammzellen durch Senkung metabolischer und proliferativer Aktivität primäre Therapieresistenz gegenüber klassischen zytostatischen Therapieverfahren sowie pharmakologischer mTOR-Inhibition.Methods: Throughout this study, the transgene model of the mouse prostate (TRAMP) expressing Simian Virus 40 t-antigens was bred and used as source for prostatic tumor tissue. Creation of single cell suspensions was performed using enzymatical tissue dissociation. Isolation of prostate cancer stem cells and intracellular immunostaining was performed using flow cytometry cell sorting (FACS). Further, well established prostate cancer cell lines of human origin were cultivated and analyzed in vitro under sterile standard cell culture conditions. Further, cell experiments were partially performed under hypoxic conditions and pharmacological blockade of the Mammalian target of rapamycin (mTOR) and Hypoxia-inducible factor 1[alpha] (HIF1A/HIF1[alpha]). Experiments performed included classical nucleomic as well as proteomic approaches such as western blot or polymerase chain reaction (PCR). Cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Spheroid formation assays were performed using matrigel and manual counting using a microscope. Viability of cells was assessed using AnnexinV stainings and FACS. Xenograft experiments were performed in NOD scid gamma (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ) mice. All animal experiments were carried out in accordance to requirements set by our institutional animal ethics committee. Results Basal CSC populations isolated from primary TRAMP tumors as well as human and murine cell lines using FACS were investigated for upregulation of known cancer promoting pathways. HIF1[alpha] was found to be overly expressed in CSCs populations when compared to their non-CSC counterparts. Further, HIF target-gene expression was abundant in hypoxic CSCs, and upregulation of protein kinase B (AKT) activity occurred through a molecular mechanism consisting of an mTOR/S6K/IRS-1 feedback loop, directly promoting survival of CSCs via deactivation of AKT feedback inhibition. Further, resistance of prostate CSCs to selective mTOR inhibitors was seen because of HIF1[alpha] upregulation. Conclusion Looking at our results, we present a novel mechanism of prostate cancer stem cell maintenance under hypoxic conditions through deregulation of the PI3K/AKT/mTOR pathway via HIF1[alpha]. We further propose the newly discovered process to be vital for CSC quiescence and maintenance by reducing CSC metabolism and growth via mTOR and supporting survival by AKT. Also, we suggest a primary drug resistance of prostate CSCs towards mTOR inhibition as well as classical cytostatic therapies.submitted by Maximilian Marhold, MDZusammenfassung in deutscher SpracheMedizinische Universität Wien, Dissertation, 2016OeBB(VLID)161614

Current Developments in Cellular Therapy for Castration Resistant Prostate Cancer: A Systematic Review of Clinical Studies

Recently, the development of immunotherapies such as cellular therapy, monoclonal antibodies, vaccines and immunomodulators has revolutionized the treatment of various cancer entities. In order to close the existing gaps in knowledge about cellular immunotherapy, specifically focusing on the chimeric antigen receptors (CAR) T-cells, their benefits and application in clinical settings, we conducted a comprehensive systematic review. Two co-authors independently searched the literature and characterized the results. Out of 183 records, 26 were considered eligible. This review provides an overview of the cellular immunotherapy landscape in treating prostate cancer, honing in on the challenges of employing CAR T-cell therapy. CAR T-cell therapy is a promising avenue for research due to the presence of an array of different tumor specific antigens. In prostate cancer, the complex microenvironment of the tumor vastly contributes to the success or failure of immunotherapies

Wiener Medizinische Wochenschrift / Whole body vibration therapy on a treatment bed as additional means to treat postprostatectomy urinary incontinence

An innovative form of whole body vibration therapy on a treatment bed (Evocell®) to fight against the disabling and isolating symptom of postoperative incontinence in a prostate cancer patient is presented. A supervised program with outpatient active pelvic floor training and a novel form of synchronous high-intensity whole body vibration therapy using the Evocell® device was performed in a patient with postprostatectomy stress urinary incontinence. The patient had previously failed regular pelvic floor exercise. During the intervention, namely a whole body vibration treatment in a lying position on a treatment bed, the patient performed active and passive pelvic floor exercises under professional guidance. Over a period of 6 weeks after starting treatment, the patient regained continence (usage of 1 safety pad). Furthermore, his ability to work increased (return to work) and his ability to attend social activities improved.(VLID)353193

Synthetic lethality guiding selection of drug combinations in ovarian cancer.

BACKGROUND:Synthetic lethality describes a relationship between two genes where single loss of either gene does not trigger significant impact on cell viability, but simultaneous loss of both gene functions results in lethality. Targeting synthetic lethal interactions with drug combinations promises increased efficacy in tumor therapy. MATERIALS AND METHODS:We established a set of synthetic lethal interactions using publicly available data from yeast screens which were mapped to their respective human orthologs using information from orthology databases. This set of experimental synthetic lethal interactions was complemented by a set of predicted synthetic lethal interactions based on a set of protein meta-data like e.g. molecular pathway assignment. Based on the combined set, we evaluated drug combinations used in late stage clinical development (clinical phase III and IV trials) or already in clinical use for ovarian cancer with respect to their effect on synthetic lethal interactions. We furthermore identified a set of drug combinations currently not being tested in late stage ovarian cancer clinical trials that however have impact on synthetic lethal interactions thus being worth of further investigations regarding their therapeutic potential in ovarian cancer. RESULTS:Twelve of the tested drug combinations addressed a synthetic lethal interaction with the anti-VEGF inhibitor bevacizumab in combination with paclitaxel being the most studied drug combination addressing the synthetic lethal pair between VEGFA and BCL2. The set of 84 predicted drug combinations for example holds the combination of the PARP inhibitor olaparib and paclitaxel, which showed efficacy in phase II clinical studies. CONCLUSION:A set of drug combinations currently not tested in late stage ovarian cancer clinical trials was identified having impact on synthetic lethal interactions thus being worth of further investigations regarding their therapeutic potential in ovarian cancer

MALAT1 Fusions and Basal Cells Contribute to Primary Resistance against Androgen Receptor Inhibition in TRAMP Mice

Targeting testosterone signaling through androgen deprivation therapy (ADT) or antiandrogen treatment is the standard of care for advanced prostate cancer (PCa). Although the large majority of patients initially respond to ADT and/or androgen receptor (AR) blockade, most patients suffering from advanced PCa will experience disease progression. We sought to investigate drivers of primary resistance against antiandrogen treatment in the TRAMP mouse model, an SV-40 t-antigen driven model exhibiting aggressive variants of prostate cancer, castration resistance, and neuroendocrine differentiation upon antihormonal treatment. We isolated primary tumor cell suspensions from adult male TRAMP mice and subjected them to organoid culture. Basal and non-basal cell populations were characterized by RNA sequencing, Western blotting, and quantitative real-time PCR. Furthermore, effects of androgen withdrawal and enzalutamide treatment were studied. Basal and luminal TRAMP cells exhibited distinct molecular signatures and gave rise to organoids with distinct phenotypes. TRAMP cells exhibited primary resistance against antiandrogen treatment. This was more pronounced in basal cell-derived TRAMP organoids when compared to luminal cell-derived organoids. Furthermore, we found MALAT1 gene fusions to be drivers of antiandrogen resistance in TRAMP mice through regulation of AR. Summarizing, TRAMP tumor cells exhibited primary resistance towards androgen inhibition enhanced through basal cell function and MALAT1 gene fusions