Reliable melanoma discrimination system offers grounds of the decision

研究成果の概要 (和文) : メラノーマの自動診断ならびに、診断根拠の客観的な提示を実現することを主目的に、3年間研究に取り組んだ。我々はメラノーマ診断の臨床で用いられるABCDルールおよび7-point checklistの全項目について、皮膚科医と同等の推定精度を得るモデルを実現した。また近年の深層学習技術を応用し、学習時間を大幅に削減しつつ熟練と皮膚科医と同等以上の識別精度を実現した。研究成果の概要 (英文) : We have investigated to build a reliable melanoma discrimination system capable of providing grounds for the decision. Our model successfully estimates all items defined in commonly used clinical findings for melanoma, i.e. a total 15 items defined in ABCD rule and 7-point checklist. We also developed efficient pre-processing techniques for melanoma discrimination for deep learning approach. Our method reduced time-consuming training time to less than 1/9, but it attained superior performance in discriminating melanomas to expert dermatologists

Application of artificial vision algorithms to images of microscopy and spectroscopy for the improvement of cancer diagnosis

El diagnóstico final de la mayoría de tipos de cáncer lo realiza un médico experto en anatomía patológica que examina muestras tisulares o celulares sospechosas extraídas del paciente. Actualmente, esta evaluación depende en gran medida de la experiencia del médico y se lleva a cabo de forma cualitativa mediante técnicas de imagen tradicionales como la microscopía óptica. Esta tarea tediosa está sujeta a altos grados de subjetividad y da lugar a niveles de discordancia inadecuados entre diferentes patólogos, especialmente en las primeras etapas de desarrollo del cáncer. La espectroscopía infrarroja por Transformada de Fourier (siglas FTIR en inglés) es una tecnología ampliamente utilizada en la industria que recientemente ha demostrado una capacidad creciente para mejorar el diagnóstico de diferentes tipos de cáncer. Esta técnica aprovecha las propiedades del infrarrojo medio para excitar los modos vibratorios de los enlaces químicos que forman las muestras biológicas. La principal señal generada consiste en un espectro de absorción que informa sobre la composición química de la muestra iluminada. Los microespectrómetros FTIR modernos, compuestos por complejos componentes ópticos y detectores matriciales de alta sensibilidad, permiten capturar en un laboratorio de investigación común imágenes hiperespectrales de alta calidad que aúnan información química y espacial. Las imágenes FTIR son estructuras de datos ricas en información que se pueden analizar individualmente o junto con otras modalidades de imagen para realizar diagnósticos patológicos objetivos. Por lo tanto, esta técnica de imagen emergente alberga un alto potencial para mejorar la detección y la graduación del riesgo del paciente en el cribado y vigilancia de cáncer. Esta tesis estudia e implementa diferentes metodologías y algoritmos de los campos interrelacionados de procesamiento de imagen, visión por ordenador, aprendizaje automático, reconocimiento de patrones, análisis multivariante y quimiometría para el procesamiento y análisis de imágenes hiperespectrales FTIR. Estas imágenes se capturaron con un moderno microscopio FTIR de laboratorio a partir de muestras de tejidos y células afectadas por cáncer colorrectal y de piel, las cuales se prepararon siguiendo protocolos alineados con la práctica clínica actual. Los conceptos más relevantes de la espectroscopía FTIR se investigan profundamente, ya que deben ser comprendidos y tenidos en cuenta para llevar a cabo una correcta interpretación y tratamiento de sus señales especiales. En particular, se revisan y analizan diferentes factores fisicoquímicos que influyen en las mediciones espectroscópicas en el caso particular de muestras biológicas y pueden afectar críticamente su análisis posterior. Todos estos conceptos y estudios preliminares entran en juego en dos aplicaciones principales. La primera aplicación aborda el problema del registro o alineación de imágenes hiperespectrales FTIR con imágenes en color adquiridas con microscopios tradicionales. El objetivo es fusionar la información espacial de distintas muestras de tejido medidas con esas dos modalidades de imagen y centrar la discriminación en las regiones seleccionadas por los patólogos, las cuales se consideran más relevantes para el diagnóstico de cáncer colorrectal. En la segunda aplicación, la espectroscopía FTIR se lleva a sus límites de detección para el estudio de las entidades biomédicas más pequeñas. El objetivo es evaluar las capacidades de las señales FTIR para discriminar de manera fiable diferentes tipos de células de piel que contienen fenotipos malignos. Los estudios desarrollados contribuyen a la mejora de métodos de decisión objetivos que ayuden al patólogo en el diagnóstico final del cáncer. Además, revelan las limitaciones de los protocolos actuales y los problemas intrínsecos de la tecnología FTIR moderna, que deberían abordarse para permitThe final diagnosis of most types of cancers is performed by an expert clinician in anatomical pathology who examines suspicious tissue or cell samples extracted from the patient. Currently, this assessment largely relies on the experience of the clinician and is accomplished in a qualitative manner by means of traditional imaging techniques, such as optical microscopy. This tedious task is subject to high degrees of subjectivity and gives rise to suboptimal levels of discordance between different pathologists, especially in early stages of cancer development. Fourier Transform infrared (FTIR) spectroscopy is a technology widely used in industry that has recently shown an increasing capability to improve the diagnosis of different types of cancer. This technique takes advantage of the ability of mid-infrared light to excite the vibrational modes of the chemical bonds that form the biological samples. The main generated signal consists of an absorption spectrum that informs of the chemical composition of the illuminated specimen. Modern FTIR microspectrometers, composed of complex optical components and high-sensitive array detectors, allow the acquisition of high-quality hyperspectral images with spatially-resolved chemical information in a common research laboratory. FTIR images are information-rich data structures that can be analysed alone or together with other imaging modalities to provide objective pathological diagnoses. Hence, this emerging imaging technique presents a high potential to improve the detection and risk stratification in cancer screening and surveillance. This thesis studies and implements different methodologies and algorithms from the related fields of image processing, computer vision, machine learning, pattern recognition, multivariate analysis and chemometrics for the processing and analysis of FTIR hyperspectral images. Those images were acquired with a modern benchtop FTIR microspectrometer from tissue and cell samples affected by colorectal and skin cancer, which were prepared by following protocols close to the current clinical practise. The most relevant concepts of FTIR spectroscopy are thoroughly investigated, which ought to be understood and considered to perform a correct interpretation and treatment of its special signals. In particular, different physicochemical factors are reviewed and analysed, which influence the spectroscopic measurements for the particular case of biological samples and can critically affect their later analysis. All these knowledge and preliminary studies come into play in two main applications. The first application tackles the problem of registration or alignment of FTIR hyperspectral images with colour images acquired with traditional microscopes. The aim is to fuse the spatial information of distinct tissue samples measured by those two imaging modalities and focus the discrimination on regions selected by the pathologists, which are meant to be the most relevant areas for the diagnosis of colorectal cancer. In the second application, FTIR spectroscopy is pushed to their limits of detection for the study of the smallest biomedical entities. The aim is to assess the capabilities of FTIR signals to reliably discriminate different types of skin cells containing malignant phenotypes. The developed studies contribute to the improvement of objective decision methods to support the pathologist in the final diagnosis of cancer. In addition, they reveal the limitations of current protocols and intrinsic problems of modern FTIR technology, which should be tackled in order to enable its transference to anatomical pathology laboratories in the future.El diagnòstic final de la majoria de tipus de càncer ho realitza un metge expert en anatomia patològica que examina mostres tissulars o cel¿lulars sospitoses extretes del pacient. Actualment, aquesta avaluació depèn en gran part de l'experiència del metge i es porta a terme de forma qualitativa mitjançant tècniques d'imatge tradicionals com la microscòpia òptica. Aquesta tasca tediosa està subjecta a alts graus de subjectivitat i dóna lloc a nivells de discordança inadequats entre diferents patòlegs, especialment en les primeres etapes de desenvolupament del càncer. L'espectroscòpia infraroja per Transformada de Fourier (sigles FTIR en anglès) és una tecnologia àmpliament utilitzada en la indústria que recentment ha demostrat una capacitat creixent per millorar el diagnòstic de diferents tipus de càncer. Aquesta tècnica aprofita les propietats de l'infraroig mitjà per excitar els modes vibratoris dels enllaços químics que formen les mostres biològiques. El principal senyal generat consisteix en un espectre d'absorció que informa sobre la composició química de la mostra il¿luminada. Els microespectrómetres FTIR moderns, compostos per complexos components òptics i detectors matricials d'alta sensibilitat, permeten capturar en un laboratori d'investigació comú imatges hiperespectrals d'alta qualitat que uneixen informació química i espacial. Les imatges FTIR són estructures de dades riques en informació que es poden analitzar individualment o juntament amb altres modalitats d'imatge per a realitzar diagnòstics patològics objectius. Per tant, aquesta tècnica d'imatge emergent té un alt potencial per a millorar la detecció i la graduació del risc del pacient en el cribratge i vigilància de càncer. Aquesta tesi estudia i implementa diferents metodologies i algoritmes dels camps interrelacionats de processament d'imatge, visió per ordinador, aprenentatge automàtic, reconeixement de patrons, anàlisi multivariant i quimiometria per al processament i anàlisi d'imatges hiperespectrals FTIR. Aquestes imatges es van capturar amb un modern microscopi FTIR de laboratori a partir de mostres de teixits i cèl¿lules afectades per càncer colorectal i de pell, les quals es van preparar seguint protocols alineats amb la pràctica clínica actual. Els conceptes més rellevants de l'espectroscòpia FTIR s'investiguen profundament, ja que han de ser compresos i tinguts en compte per dur a terme una correcta interpretació i tractament dels seus senyals especials. En particular, es revisen i analitzen diferents factors fisicoquímics que influeixen en els mesuraments espectroscòpiques en el cas particular de mostres biològiques i poden afectar críticament la seua anàlisi posterior. Tots aquests conceptes i estudis preliminars entren en joc en dues aplicacions principals. La primera aplicació aborda el problema del registre o alineació d'imatges hiperespectrals FTIR amb imatges en color adquirides amb microscopis tradicionals. L'objectiu és fusionar la informació espacial de diferents mostres de teixit mesurades amb aquestes dues modalitats d'imatge i centrar la discriminació en les regions seleccionades pels patòlegs, les quals es consideren més rellevants per al diagnòstic de càncer colorectal. En la segona aplicació, l'espectroscòpia FTIR es porta als seus límits de detecció per a l'estudi de les entitats biomèdiques més xicotetes. L'objectiu és avaluar les capacitats dels senyals FTIR per discriminar de manera fiable diferents tipus de cèl¿lules de pell que contenen fenotips malignes. Els estudis desenvolupats contribueixen a la millora de mètodes de decisió objectius que ajuden el patòleg en el diagnòstic final del càncer. A més, revelen les limitacions dels protocols actuals i els problemes intrínsecs de la tecnologia FTIR moderna, que haurien d'abordar per permetre la seva transferència als laboratoris d'anatomia patològica en el futur.Peñaranda Gómez, FJ. (2018). Application of artificial vision algorithms to images of microscopy and spectroscopy for the improvement of cancer diagnosis [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/99748TESI

VIBRATIONAL SPECTROSCOPY FOR THE ASSESSMENT OF VULVAL DISEASE

Vibrational spectroscopic diagnostic techniques have significant potential to improve the care of women with benign, premalignant and malignant vulval diseases by reducing the reliance on traditional biopsy and histopathology. These techniques also have the potential to augment clinicians’ ability to differentiate different types of vulval disease at the time of surgery for neoplastic vulval disease. In addition, vibrational spectroscopic techniques offer the opportunity to assess molecular changes associated with the development of vulval cancer that are not apparent on routine histopathological assessment. The work outlined in this thesis evaluates the role of emerging techniques in vibrational spectroscopy to address this need within three key themes: 1. Developmentofavibrationalspectroscopicdiagnostictechniquetoreducethe reliance on traditional biopsy and histopathological diagnosis. 2. Developmentofavibrationalspectroscopicdiagnostictechniqueforimproving the delineation of disease margins at the time of surgery for pre-malignant and malignant vulval conditions. 3. Evaluation of a vibrational spectroscopic tool for augmenting and automating aspects of vulval histopathology. Raman spectroscopic mapping of 91 fresh frozen vulval tissue sections combined with multivariate spectral analysis was used to demonstrate that malignant vulval disease could be differentiated from non-neoplastic and premalignant vulval disease with a sensitivity of 97% and specificity of 78%. The technique was then tested in experimental conditions closer to in-vivo application, measuring spectra from 91 whole fresh frozen tissue blocks using microscope and probe Raman systems. This demonstrated the technique could differentiate malignant from non-neoplastic and premalignant vulval disease with sensitivities of 84% to 92% and specificities of 84% to 64% respectively. In a separate investigation vulval tissue blocks from 27 women with suspected lichen sclerosus underwent Raman spectroscopic point measurements. Multivariate analysis demonstrated Raman spectroscopy could be used to differentiate lichen sclerosus from other vulval disorders with a similar clinical appearance with a sensitivity of sensitivity of 91% and specificity of 80%. Fourier transform infrared (FTIR) spectroscopic mapping of 93 fixed paraffin embedded tissue sections was used to demonstrate that malignant vulval disease could be differentiated from non-neoplastic and premalignant with vulval disease with an approximate sensitivity of 100% and specificity of 79%. In addition FTIR spectroscopy was used to differentiate molecular changes in vulval intraepithelial neoplasia (VIN) and lichen sclerosus (LS) found in association with vulval squamous cell carcinoma (SCC). Analysis of FTIR spectroscopic tissue maps from 48 patients demonstrated the technique could differentiate LS associated with SCC with a sensitivity of approximately 100% and specificity of 84% and VIN associated with SCC with a sensitivity of approximately 100% and specificity 58%. This thesis demonstrates the considerable potential of vibrational spectroscopy in this clinical setting. The research has made significant progress in each of the three themes outlined above and indicates that further work is warranted to develop the techniques towards routine clinical application

Improving biomarker assessment in breast pathology

The accuracy of prognostic and therapy-predictive biomarker assessment in breast tumours is crucial for management and therapy decision in patients with breast cancer. In this thesis, biomarkers used in clinical practice with emphasise on Ki67 and HER2 were studied using several methods including immunocytochemistry, in situ hybridisation, gene expression assays and digital image analysis, with the overall aim to improve routine biomarker evaluation and clarify the prognostic potential in early breast cancer. In paper I, we reported discordances in biomarker status from aspiration cytology and paired surgical specimens from breast tumours. The limited prognostic potential of immunocytochemistry-based Ki67 scoring demonstrated that immunohistochemistry on resected specimens is the superior method for Ki67 evaluation. In addition, neither of the methods were sufficient to predict molecular subtype. Following this in paper II, biomarker agreement between core needle biopsies and subsequent specimens was investigated, both in the adjuvant and neoadjuvant setting. Discordances in Ki67 and HER2 status between core biopsies and paired specimens suggested that these biomarkers should be re-tested on all surgical breast cancer specimens. In paper III, digital image analysis using a virtual double staining software was used to compare methods for assessment of proliferative activity, including mitotic counts, Ki67 and the alternative marker PHH3, in different tumour regions (hot spot, invasive edge and whole section). Digital image analysis using virtual double staining of hot spot Ki67 outperformed the alternative markers of proliferation, especially in discriminating luminal B from luminal A tumours. Replacing mitosis in histological grade with hot spot-scored Ki67 added significant prognostic information. Following these findings, the optimal definition of a hot spot for Ki67 scoring using virtual double staining in relation to molecular subtype and outcome was investigated in paper IV. With the growing evidence of global scoring as a superior method to improve reproducibility of Ki67 scoring, a different digital image analysis software (QuPath) was also used for comparison. Altogether, we found that automated global scoring of Ki67 using QuPath had independent prognostic potential compared to even the best virtual double staining hot spot algorithm, and is also a practical method for routine Ki67 scoring in breast pathology. In paper V, the clinical value of HER2 status was investigated in a unique trastuzumab-treated HER2-positive cohort, on the protein, mRNA and DNA levels. The results demonstrated that low levels of ERBB2 mRNA but neither HER2 copy numbers, HER2 ratio nor ER status, was associated with risk of recurrence among anti-HER2 treated breast cancer patients. In conclusion, we have identified important clinical aspects of Ki67 and HER2 evaluation and provided methods to improve the prognostic potential of Ki67 using digital image analysis. In addition to protein expression of routine biomarkers, mRNA levels by targeted gene expression assays may add further prognostic value in early breast cance

Aprendizado profundo em triagem de melanoma

Orientadores: Eduardo Alves do Valle Junior, Lin Tzy LiDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Elétrica e de ComputaçãoResumo: De todos os cânceres de pele, melanoma representa apenas 1% dos casos, mas 75% das mortes. O prognóstico do melanoma é bom quando detectado cedo, mas deteriora rápido ao longo que a doença progride. Ferramentas automatizadas podem prover triagem mais rápida, ajudando médicos a focar em pacientes ou lesões de risco. As características da doença --- raridade, letalidade, rápida progressão, e diagnóstico sutil --- fazem a triagem de melanoma automática particularmente desafiadora. O objetivo deste trabalho é melhor compreender como Deep Learning pode ser utilizado --- mais precisamente, Redes Neurais Convolucionais --- para classificar corretamente imagens de lesões de pele. Para isso, este trabalho está dividido em duas linhas de pesquisa. Primeiro, o estudo está focado na transferibilidade de características das redes CNN pré-treinadas. O objetivo principal desse tópico é estudar como as características transferidas se comportam em diferentes esquemas, com o objetivo de gerar melhores características para a camada de decisão. Em um segundo tópico, esse estudo incidirá na melhoria das métricas de classificação, que é o objetivo geral. Sobre a transferibilidade das características, foram realizados experimentos para analisar a forma como os diferentes esquemas de transferência afetariam a Área sob a Curva ROC (AUC): treinar uma CNN a partir do zero; transferir o conhecimento de uma CNN pré-treinada com imagens gerais ou específicas; realizar uma transferência dupla, que é uma sequência de treinamento onde em um primeiro momento a rede é treinada com imagens gerais, em um segundo momento com as imagens específicas, e, finalmente, em um terceiro momento com as imagens de melanoma. A partir desses experimentos, aprendemos que a transferência de aprendizagem é uma boa prática, assim como é o ajuste fino. Os resultados também sugerem que modelos mais profundos conduzem a melhores resultados. Hipotetizamos que a transferência de aprendizagem de uma tarefa relacionada sob ponto de vista médico (no caso, a partir de um dataset de imagens de retinopatia) levaria a melhores resultados, especialmente no esquema de transferência dupla, mas os resultados mostraram o oposto, sugerindo que a adaptação de tarefas muito específicas representa desafios específicos. Sobre a melhoria das métricas, discute-se o pipeline vencedor utilizado no International Skin Imaging Collaboration (ISIC) Challenge 2017, alcançando o estado da arte na classificação de melanoma com 87.4% AUC. A solução é baseada em stacking/meta learning dos modelos Inception v4 e Resnet101, realizando fine tuning enquanto executa a aumentação de dados nos conjuntos de treino e teste. Também comparamos diferentes técnicas de segmentação --- multiplicação elemento a elemento da imagem da lesão de pele e sua máscara de segmentação, e utilizar a máscara de segmentação como quarto canal --- com uma rede treinada sem segmentação. A rede sem segmentação é a que obteve melhor desemepnho (96.0% AUC) contra a máscara de segmentação como quarto canal (94.5% AUC). Nós também disponibilizamos uma implementação de referência reprodutível com todo o código desenvolvido para as contribuições desta dissertaçãoAbstract: From all skin cancers, melanoma represents just 1% of cases, but 75% of deaths. Melanoma¿s prognosis is good when detected early, but deteriorates fast as the disease progresses. Automated tools may play an essential role in providing timely screening, helping doctors focus on patients or lesions at risk. However, due to the disease¿s characteristics --- rarity, lethality, fast progression, and diagnosis subtlety --- automated screening for melanoma is particularly challenging. The objective of this work is to understand better how can we use Deep Learning --- more precisely, Convolutional Neural Networks --- to correctly classify images of skin lesions. This work is divided into two lines of investigation to achieve the objective. First, the study is focused on the transferability of features from pretrained CNN networks. The primary objective of that thread is to study how the transferred features behave in different schemas, aiming at generating better features to the classifier layer. Second, this study will also improve the classification metrics, which is the overall objective of this line of research. On the transferability of features, we performed experiments to analyze how different transfer schemas would impact the overall Area Under the ROC Curve (AUC) training a CNN from scratch; transferring from pretrained CNN on general and specific image databases; performing a double transfer, in a sequence from general to specific and finally melanoma databases. From those experiments, we learned that transfer learning is a good practice, as is fine tuning. The results also suggest that deeper models lead to better results. We expected that transfer learning from a related task (in the case, from a retinopathy image database) would lead to better outcomes, but results showed the opposite, suggesting that adaptation from particular tasks poses specific challenges. On the improvement of metrics, we discussed the winner pipeline used in the International Skin Imaging Collaboration (ISIC) Challenge 2017, reaching state-of-the-art results on melanoma classification with 87.4% AUC. The solution is based on the stacking/meta-learning from Inception v4 and Resnet101 models, fine tuning them while performing data augmentation on the train and test sets. Also, we compare different segmentation techniques - elementwise multiplication of the skin lesion image and its mask, and input the segmentation mask as a fourth channel - with a network trained without segmentation. The network with no segmentation is the one who performs better (96.0% AUC) against segmentation mask as a fourth channel (94.5% AUC). We made available a reproducible reference implementation with all developed source code for the contributions of this thesisMestradoEngenharia de ComputaçãoMestre em Engenharia Elétrica133530/2016-7CNP