160 research outputs found

    2023 Summer Experience Program Abstracts

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    https://openworks.mdanderson.org/sumexp23/1130/thumbnail.jp

    The impact of realistic environmental chemical exposure on male gonadal development and reproductive health

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    Continuing declines in human male reproductive health are of increasing concern. Many believe low-dose exposure to vast numbers of chemicals through the environment, particularly during fetal development, are a contributary factor in this decline. To address limitations with traditional, component-based methodologies of assessing chemical mixtures, this research utilised a unique, ovine based, whole-mixture exposure model. This model was used to investigate the impact of gestational exposure to realistic numbers of chemicals, at appropriately low doses, on male reproductive development. The research detailed herein characterises exposure-induced changes to the testes of neonatal, pre-pubertal, and adult male offspring of mothers exposed to an environmental chemical mixture prior to and during pregnancy. A testicular dysgenesis syndrome (TDS)-like phenotype was described in neonatal and prepubertal testes. This TDS-like phenotype was complemented by transcriptomic analyses which showed an extremely high degree of similarity between the testicular transcriptome of the affected pre-pubertal male offspring and those of human TDS patients. While this phenotype was not apparent in the same manner by adulthood, morphological and transcriptomic alterations were still apparent. This both exemplifies the potential for xenobiotic exposure during fetal development to impact reproductive health in later life, despite the cessation of exposure at birth, and indicates periods of post-partum vulnerability to xenobiotic exposure crucial to the persistence of or recovery from the TDS-like phenotype. Further investigations following transcriptomic analyses identified perturbations in the transcription, activation, and/or nuclear localisation of various transcription factors. Of these, there is supporting evidence that one (HIF1α) may have an important role in the pathogenesis of the TDS-like phenotype, while another (CREB1) may facilitate an amount of post-exposure recovery and might also be important in determining susceptibility or resistance to developing the TDS-phenotype. Overall, these findings strengthen the increasing evidence that gestational exposure to realistic levels and mixtures of environmental chemicals can have a negative impact on male reproductive health and provides leads for future investigations into the pathogenesis of TDS

    Advances in Concepts, Ideas, and Methods Relevant to Fine Needle Aspiration Biopsy of Thyroid and Cervical Lymph Node

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    With the increasingly used semi-thyroidectomy and rapid progress in ultrasound-guided thermal ablation therapy for treatment of papillary thyroid carcinoma (PTC) and cervical lymph node metastasis from PTC, ultrasound-guided fine needle aspiration biopsy (FNAB) has got the mainstream position in pre-treatment cytopathologic diagnosis of PTC. How to acquire adequate and qualified cellular specimen for cytological examination has been described in several published expert consensus and practice guidelines. However, new issues continue to emerge in the real world of thyroid FNAB practice, and most of them are rooted in the perception and skills of the physician or technician who conduct FNAB. In this chapter, a series of new concept, idea, and technical methods are to be introduced and discussed. We believe that properly addressing these issues will facilitate the better implementation of FNAB and promote the new therapeutic modalities such as the thermal ablation to better progress

    A Single-Molecule Bioelectronic Portable Array for Early Diagnosis of Pancreatic Cancer Precursors

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    A cohort of 47 patients is screened for pancreatic cancer precursors with a portable 96-well bioelectronic sensing-array for single-molecule assay in cysts fluid and blood plasma, deployable at point-of-care (POC). Pancreatic cancer precursors are mucinous cysts diagnosed with a sensitivity of at most 80% by state-of-the-art cytopathological molecular analyses (e.g., KRASmut DNA). Adding the simultaneous assay of proteins related to malignant transformation (e.g., MUC1 and CD55) is deemed essential to enhance diagnostic accuracy. The bioelectronic array proposed here, based on single-molecule-with-a-large-transistor (SiMoT) technology, can assay both nucleic acids and proteins at the single-molecule limit-of-identification (LOI) (1% of false-positives and false-negatives). It comprises an enzyme-linked immunosorbent assay (ELISA)-like 8 × 12-array organic-electronics disposable cartridge with an electrolyte-gated organic transistor sensor array, and a reusable reader, integrating a custom Si-IC chip, operating via software installed on a USB-connected smart device. The cartridge is complemented by a 3D-printed sensing gate cover plate. KRASmut, MUC1, and CD55 biomarkers either in plasma or cysts-fluid from 5 to 6 patients at a time, are multiplexed at single-molecule LOI in 1.5 h. The pancreatic cancer precursors are classified via a machine-learning analysis resulting in at least 96% diagnostic-sensitivity and 100% diagnostic-specificity. This preliminary study opens the way to POC liquid-biopsy-based early diagnosis of pancreatic-cancer precursors in plasma

    Malignant salivary gland tumours:Clinical behaviour and prognosticators

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    Digital Histopathology of Cancer

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    Syöpä on merkittävä ja yleistyvä kansansairaus. Maailman terveysjärjestön mukaan syöpä on maailmanlaajuisesti toiseksi yleisin kuolinsyy sydän- ja verisuonitautien jälkeen. Jos ei-melanoottisia ihosyöpiä ei oteta huomioon, ovat tavallisimmat syöpätyypit naisilla rintasyöpä ja miehillä keuhkosyöpä ja eturauhassyöpä. Sitä mukaa kun syövän biologisten syntymekanismien ymmärrys on lisääntynyt, ovat myös hoitovaihtoehdot lisääntyneet. Useampi kuin joka neljäs uusi lääke, joka lanseerattiin vuosina 2010-2018, oli tarkoitettu syövän hoitoon. Jotta potilas voisi hyötyä tarjolla olevasta laajasta syöpälääkevalikoimasta ja minimoida lääkkeiden haittavaikutukset, tulee hoito kohdistaa hänen yksilölliseen syöpäänsä. Tätä varten syöpä on sekä diagnosoitava luotettavasti että luokiteltava yksityiskohtaisesti. Vaikka kajoamattomat kuvantamistutkimukset kuten magneettikuvaus ovat viime vuosina kehittyneet huomattavasti, on syöpädiagnostiikan perusta edelleen histopatologiassa eli leikkauksessa tai neulanäytteenotossa poistetun kudoksen mikroskooppisessa tutkimuksessa. Valomikroskooppi on pysynyt patologin pääasiallisena työvälineenä yli puolentoista vuosisadan ajan. Se on sallinut kudoksen tarkastelun aina solutasolle saakka ja jopa sitä pienempiin rakenteisiin. Tärkeitä lisätutkimuksia tavallisen valomikroskooppisen tutkimuksen lisäksi ovat proteiiniantigeenien osoittamistutkimukset, kuten immunohistokemia ja in situ - hybridisaatio, joita voidaan käyttää syöpäkudoksen luokittelemiseen. Syövän tarkalla diagnosoimisella ja luokittelulla on haasteensa. Yksi sellainen on Suomessa ja ulkomailla vallitseva pula patologeista. Toinen haaste liittyy kasvainten välisen vaihtelun arviointiin, joka on tärkeää kasvainten kasvutaipumuksen luokittelussa (esim. eturauhassyövän Gleason-luokitus) ja tiettyjen värjäysten tulkinnassa (esim. rintasyövän HER2-värjäytyminen). Todellisen biologisen vaihtelun lisäksi vaihtelua esiintyy patologien välisissä arvioissa (interobserver variation) sekä saman patologin luokitteluissa eri ajan hetkellä (intraobserver variation). Kolmas haaste on itse valomikroskooppi. Vaikka se on luotettava, halpa ja helppokäyttöinen diagnostiikkalaite, on sillä omat puutteensa modernin patologian laboratorion työnkulussa. Digitaalihistopatologia edustaa uutta tapaa toteuttaa patologin pääasiallinen työtehtävä syöpäpotilaan hoidossa: asettaa diagnoosi ja luokitella syöpä yksityiskohtaisesti. Siirtyminen valomikroskoopista tietokoneympäristöön tarjoaa monia etuja, joista muutamia on tutkittu tässä väitöskirjassa. Tämän tutkimuksen tarkoituksena oli kehittää ja testata digitaalipatologian sovelluksia syöpädiagnostiikan parantamiseksi. Osatöissä tutkittiin eturauhassyövän Gleason-luokituksen opettamista ja standardointia, rinta- ja eturauhassyövän immunohistokemiallisten värjäysten tulkintaa, digitaalinäytteille kehitettyä kuvanpakkausmenetelmää, sekä näyteskannerin optimaalisen kuvausresoluution määrittämistä. Väitöskirjassa osoitetaan, että digitaalinäytteitä voi käyttää eturauhaskoepalan Gleason-luokituksen tekemiseen ja että internet-pohjainen ohjelma voi edistää tulkitsijoiden välisen vaihtelun määrittämistä sekä Gleason-luokituksen opettamista ja standardisointia. Gleason-luokituksen ohella toinen tärkeä osa eturauhassyövän histopatologiaa on immunohistokemiallisten värjäysten tulkinta. Tässä väitöskirjassa esitetään menetelmä, jolla kahta digitaalinäytettä voidaan tutkia yhtäaikaisesti ja synkronoidusti. Menetelmää testattiin eturauhassyövän immunohistokemiallisella AMACR–p63-kaksoisvärjäyksellä yhdessä rutiininomaisen hematoksyliini–eosiini- värjäyksen kanssa. Tutkimuksessa osoitettiin, että tekniikkaa voidaan käyttää hyväksi histopatologian opetuksessa ja valikoiduissa tapauksissa kliinisessä diagnostiikassa. Keskeinen asia rintasyövän diagnostiikassa on HER2-statuksen tutkiminen, koska kasvaimia, joissa HER2 on yli-ilmentynyt, voidaan hoitaa anti-HER2- lääkkeillä. Yhdessä osatöistä tutkittiin digitaalisen kuva-analyysin käyttöä niin valomikroskooppikuvilla kuin digitaalinäytteillä tarkoituksena auttaa patologia määrittämään kirurgisesti poistetun kasvainkudoksen HER2-status. Työssä osoitettiin, että ilmaista ja kaikille avointa ohjelmistoa käyttämällä voitiin vähentää HER2-statuksen suhteen vaikeatulkintaisten tapausten määrää. Digitaalihistopatologian käyttöönotto rutiinidiagnostiikkaan on laajentumassa nopeasti. Yksi tekninen haaste on digitaalinäytteiden vaatiman suuren tallennuskapasiteetin hallinta. Tarve tallentaa suuria määriä tietoa edellyttää digitaalinäytteiden kuvanlaadun ja tiedostokoon yhteensovittamista. Yhdessä tämän väitöskirjan osatöistä tutkittiin skannerimikroskoopin optimaalisen kuvausresoluution määrittämistä. Menetelmää voidaan hyödyntää esimerkiksi vertailtaessa skannereita ennen hankintaa. Toisessa osatyössä esiteltiin uusi kuvanpakkausmenetelmä, joka suunniteltiin varta vasten histopatologisia digitaalinäytteitä varten niiden tiedostokoon minimoimiseksi ja siten tallennuskustannusten pienentämiseksi. Tämän väitöskirjan kaksi ensimmäistä osatyötä edustavat digitaalipatologian alkutaivalta ja tutkimuskenttä on kehittynyt sittemmin, mahdollisesti pieneltä osin edellä mainittujen tutkimusten löydösten myötä. Yhteenvetona osatyöt toivottavasti vievät digitaalipatologian alaa eteenpäin ja siten edesauttavat syöpäpotilaiden hoitoa.Cancer is a significant and growing public health concern. According to the World Health Organisation's estimates it is – after cardiovascular diseases – the second leading cause of death worldwide. Excluding non-melanoma skin cancers the most common types of cancer are for women breast cancer and for men lung cancer followed by prostate cancer. While the biological understanding of cancer has expanded, so too has the selection of available treatments. More than one fourth of all new medicines entering the market during 2010-2018 were for treating cancer. In order for a patient to benefit from the wide variety of cancer treatments, and avoid adverse effects, their unique cancer has to be matched with the appropriate treatment. For this the cancer needs to be both diagnosed accurately and classified in detail. Although non-invasive imaging methods, such as magnetic resonance imaging, have evolved substantially in recent years, the basis of cancer diagnosis is still in histopathology, that is, the pathological evaluation of tissue removed through surgery or needle biopsy. The light microscope has remained the pathologist's main diagnostic tool for a century and a half allowing for the examination of tissue down to cellular – and even subcellular – level. Important adjuncts to routine histopathological staining of tissue, needed for light microscopy, are techniques allowing for the visualization of protein antigens and nucleic acid in the tissue. These techniques, among which are immunohistochemistry and in situ hybridization, respectively, can be used for instance in the molecular characterization of cancer. There are challenges in meeting the need for accurate diagnosing and characterization of cancer. One such challenge is posed by the shortage of pathologists observed in Finland and elsewhere. Another challenge is the variability in the interpretation of the tumor growth pattern (grading, such as Gleason grading in prostate cancer) and in the interpretation of certain tissue staining patterns (such as the immunohistochemical staining of the HER2 molecule in breast cancer). This variability manifests itself both between pathologists (interobserver variation) and also in the same pathologist's work over time (intraobserver variation). A third challenge is presented by the fact that the light microscope – although a reliable, cheap, and easy-to-use diagnostic tool – has shortcomings in the modern day pathology service. Digital histopathology presents a new way of carrying out the central task of a pathologist in managing cancer patients, namely making the diagnosis and characterising the tumor in detail. Making the shift from a light microscope to a computer environment offers many benefits, some of which have been examined in this dissertation. The present study was carried out with the purpose of developing and testing applications of digital pathology in order to improve the histopathological diagnosis of cancer. The individual studies looked at advancing the teaching and standardization of Gleason grading or prostate cancer, aiding in the interpretation of immunohistochemical staining of prostate and breast cancer, as well as facilitating the implementation of digital pathology by way of a novel whole slide image optimised image compression algorithm and mapping the determinants of an optimal imaging resolution for a whole slide scanner. We demonstrated that whole slide images can be used to assess the Gleason grade of a prostate biopsy and that the use of an internet based platform can be beneficial in assessing interobserver variation in the grading and teaching and standardising the grading. Besides Gleason grading another important aspect of prostate histopathology is the interpretation of immunohistochemistry. We created a method of viewing two whole slide images simultaneously and synchronously and tested this method in visualising the AMACR-p63 double stain along with normal hematoxylin and eosin staining of prostate biopsies. We showed that this technique can be used for histopathology education as well as in clinical diagnostics in selected cases. A key issue in breast cancer diagnostics is defining the HER2 status of a tumor, that is, whether the tumor overexpresses the molecule and can then be treated with HER2 antibody based drugs. We studied the use of digital image analysis, using both photomicrographs and whole slide images, in aiding the pathologist in defining the HER2 status on a breast cancer surgical resection specimen. We showed that using a free and publicly available image analysis software can help to resolve cases otherwise deemed equivocal by conventional light microscopy. The introduction of digital histopathology into routine diagnostic work is underway. One technical challenge is managing the large amounts of image data generated by whole slide images. When there is a need to store large numbers of whole slide images it is essential to strike a balance between image fidelity and file size. To deal with this issue we studied the optimal imaging resolution of a whole slide scanner using a methodology that can be utilised for instance in comparing whole slide scanners before acquiring one. In addition we introduced a novel way of image compression suited for whole slide images in order to reduce the storage footprint, and cost, of whole slide images. The first two studies in this dissertation represent the very beginnings of whole slide imaging in pathology, and the field has advanced since then, perhaps in small part due to the findings in these studies. Taken together, the findings in this dissertation can hopefully advance the use of digital pathology in cancer diagnostics and thereby improve the care of cancer patients

    The effect of epigenetic therapies on Glioblastoma Multiforme cancer stem cells

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    Glioblastoma Multiforme (GBM) is one of the most aggressive types of brain tumour; it is hard to treat with conventional therapy and often shows tumour regrowth after surgical removal. Recent studies have indicated the presence of cancer stem cells in GBM tumours, which are more resistant to therapy and driver of tumour recurrence after treatment. Here, we investigate the effect of several epigenetic inhibitors on two GBM stem cell lines, G7 and E2. Cells were not sensitive to the BET inhibitor OTX015, but the EZH2 inhibitors EPZ6438, UNC1999 and GSK343 all significantly reduced colony formation. The three EZH2 inhibitors induced expression of senescence-associated β-galactosidase, sensitised both cell lines to the alkylating agent temozolomide, and sensitised the E2 cell line to radiation. RNA-seq analysis following treatment with EZH2 inhibitors revealed a significant induction of genes related to neurogenesis and neuronal function, particular in the G7 cell line. ChIP-seq analysis of the epigenetic changes following EZH2 treatment was used to identify genes that were both upregulated and showed a decrease in histone H3 lysine 27 tri-methylation, which is the epigenetic mark deposited by EZH2. Several regulatory factors were identified that are potential candidates for driving the neuronal differentiation of the cancer stem cells. One of these candidates is the transcription factor PAX2, which is upregulated in both G7 and E2 cells. PAX2 is essential for the correct development of the inner ear and the eye, and is known to regulate expression of the neural transcription factors NEUROG1, NEUROD1 and ATOH1. Based on these data, it is hypothesise that the EZH2 inhibitors reduce GBM stem cell proliferation by inducing neuronal differentiation of the cancer stem cells, in addition to causing therapy induced senescence. Further work is required to investigate whether the candidate regulatory genes that were identified, including PAX2, do indeed play a role in driving this differentiation. Sensitisation of the GBM stem cells to temozolomide and radiation is a promising line of enquiry for further investigation into improving treatments for GBM
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