Background. First Study: Fluorescence confocal microscopy (FCM) enables real time image acquisition of excised tissue at high resolution in the assessment of micrographic controlled surgery margins. The new generation FCM incorporates reflectance and fluorescence modalities separately or simultaneously, in green and white or hematoxylin and eosin like (H&E-like). Second Study: actinic keratosis (AKs) are precancerous lesions that can progress to squamous cell carcinoma (SCC). The progression from AK to SCC was proposed to occur following a progressive multi-step pathway. Recently AKs with atypical cells limited to the basal layer (AKs I) have been demonstrated to be the most common precursors of SCCs arising from a differentiated pathway. Objectives. First Study: assessing the diagnostic accuracy of FCM modalities and the evaluators compared to gold standard. Second study: assessing the complete metabolomic fingerprint of AKI, II, III and SCC by HR-MAS-NMR spectroscopy in order to evaluate the hypothesis of the grade-independent progression. Progression from AKs to SCCs was evaluated also using histopathology. Methods. First study: retrospective analysis of basal cell carcinoma treated with micrographic controlled surgery. FCM images of tumor margins were assessed by pathologists and dermatologists. Evaluator agreement was assessed with Cohen’s kappa and the diagnostic performance of all evaluators with the ROC curve and the AUC. Second study: metabolomic data were obtained through HR-MAS NMR spectroscopy. The spectral profiles were analyzed through multivariate statistical analysis using MetaboAnalyst 5.0. Histological examination was performed on hematoxylin and eosin sections, statistical analysis was performed using STATA® software version 14 . Results. First study: H&E-like modality achieved the best diagnostic accuracy, confirming 94% overall correct diagnoses. At ROC curve the highest AUC was associated with the H&E-like mode, reporting the highest sensitivity of 83.3% and a specificity of 96.1%. Second study: SCCs and AKI were significantly associated in terms of metabolite content. In the logistic regression model, the presence of parakeratosis in AKs appeared to be less frequently associate with SCCs, while AKs with hypertrophy had a higher risk of being associated with SCC. Conclusions. First study suggests that FCM H&E-like images are associated with the highest diagnostic accuracy, interpreted by both dermatologists and pathologists. The future application of FCM in micrographic controlled surgery may decrease the number of surgical stages, reducing surgical time and related costs. Second study: all the findings, derived from metabolomics and histopathology, support that AKI are different from healthy skin and share features with SCC, implicating that all AKs should be treated because it is not possible to predict their evolution to SCC.

Background. Primo studio: la microscopia confocale a fluorescenza (FCM) consente l'acquisizione di immagini in tempo reale ad alta risoluzione del tessuto asportato per la valutazione dei margini tumorali della chirurgia microscopicamente controllata. L'FCM di nuova generazione incorpora modalità di riflettanza e fluorescenza separatamente o simultaneamente (BW), in verde e bianco (GW) o come ematossilina ed eosina (H&E-like). Secondo studio: le cheratosi attiniche (AK) sono lesioni precancerose che possono evolvere in carcinoma squamocellulare (SCC). In passato si ipotizzava che la progressione da AK a SCC secondo un progressivo percorso a più stadi. Recentemente è stato dimostrato che gli AK con cellule atipiche limitate allo strato basale (AK I) sono i precursori più comuni degli SCC, secondo un differentiated pathway. Obiettivi. Primo studio: valutazione dell'accuratezza diagnostica delle modalità FCM e dei valutatori rispetto al gold standard. Secondo studio: valutazione dei profili metabolomici di AKI, II, III e SCC mediante spettroscopia HR-MAS-NMR al fine di valutare l'ipotesi della progressione indipendente dal grado. La progressione da AK a SCC è stata valutata anche mediante l'istopatologia. Metodi. Primo studio: analisi retrospettiva di carcinomi basocellulari trattati con chirurgia microscopicamente controllata. Le immagini FCM dei margini dei tumori sono state valutate da 2 patologi e 2 dermatologi. La concordanza dei valutatori è stata valutata con la kappa di Cohen e le prestazioni diagnostiche di tutti i valutatori con la curva ROC e l'AUC. Secondo studio: i dati metabolomici sono stati ottenuti mediante spettroscopia HR-MAS NMR. I profili degli spettri sono stati analizzati mediante analisi statistica multivariata utilizzando MetaboAnalyst 5.0. L'esame istologico è stato eseguito su sezioni di ematossilina ed eosina, l'analisi statistica è stata eseguita utilizzando il software STATA® versione 14 . Risultati. Primo studio: la modalità H&E-like ha ottenuto la migliore accuratezza diagnostica, confermando il 94% di diagnosi complessivamente corrette. Alla curva ROC l'AUC più alta era associata alla modalità H&E-like, riportando la sensibilità più alta dell'83,3% e una specificità del 96,1%. Secondo studio: SCC e AKI erano significativamente simili in termini di contenuto di metaboliti. Nel modello di regressione logistica, la presenza di paracheratosi negli AK sembrava essere associata meno frequentemente agli SCC, mentre gli AK con ipertrofia avevano un rischio maggiore di essere associati a SCC. Conclusioni. Il primo studio suggerisce che le immagini FCM H&E-like presentano la migliore accuratezza diagnostica, interpretata sia dai dermatologi che dai patologi. L'applicazione dell' FCM nella chirurgia microscopicamente controllata potrebbe diminuire il numero di stadi chirurgici, riducendo i tempi dell'intervento e i relativi costi. Secondo studio: tutti i risultati, derivati dalle analisi di metabolomica e di istopatologia, supportano che l'AKI è diverso dalla cute sana e condivide alcune caratteristiche con l'SCC, implicando che tutti gli AK dovrebbero essere trattati, indipendentemente dal grado, perché non è possibile predire la loro evoluzione a SCC.

Nuove tecnologie per la diagnosi e il trattamento dei non melanoma skin cancers / Camilla Reggiani , 2023 May 19. 34. ciclo, Anno Accademico 2020/2021.

Nuove tecnologie per la diagnosi e il trattamento dei non melanoma skin cancers

Reggiani, Camilla
2023

Abstract

Background. First Study: Fluorescence confocal microscopy (FCM) enables real time image acquisition of excised tissue at high resolution in the assessment of micrographic controlled surgery margins. The new generation FCM incorporates reflectance and fluorescence modalities separately or simultaneously, in green and white or hematoxylin and eosin like (H&E-like). Second Study: actinic keratosis (AKs) are precancerous lesions that can progress to squamous cell carcinoma (SCC). The progression from AK to SCC was proposed to occur following a progressive multi-step pathway. Recently AKs with atypical cells limited to the basal layer (AKs I) have been demonstrated to be the most common precursors of SCCs arising from a differentiated pathway. Objectives. First Study: assessing the diagnostic accuracy of FCM modalities and the evaluators compared to gold standard. Second study: assessing the complete metabolomic fingerprint of AKI, II, III and SCC by HR-MAS-NMR spectroscopy in order to evaluate the hypothesis of the grade-independent progression. Progression from AKs to SCCs was evaluated also using histopathology. Methods. First study: retrospective analysis of basal cell carcinoma treated with micrographic controlled surgery. FCM images of tumor margins were assessed by pathologists and dermatologists. Evaluator agreement was assessed with Cohen’s kappa and the diagnostic performance of all evaluators with the ROC curve and the AUC. Second study: metabolomic data were obtained through HR-MAS NMR spectroscopy. The spectral profiles were analyzed through multivariate statistical analysis using MetaboAnalyst 5.0. Histological examination was performed on hematoxylin and eosin sections, statistical analysis was performed using STATA® software version 14 . Results. First study: H&E-like modality achieved the best diagnostic accuracy, confirming 94% overall correct diagnoses. At ROC curve the highest AUC was associated with the H&E-like mode, reporting the highest sensitivity of 83.3% and a specificity of 96.1%. Second study: SCCs and AKI were significantly associated in terms of metabolite content. In the logistic regression model, the presence of parakeratosis in AKs appeared to be less frequently associate with SCCs, while AKs with hypertrophy had a higher risk of being associated with SCC. Conclusions. First study suggests that FCM H&E-like images are associated with the highest diagnostic accuracy, interpreted by both dermatologists and pathologists. The future application of FCM in micrographic controlled surgery may decrease the number of surgical stages, reducing surgical time and related costs. Second study: all the findings, derived from metabolomics and histopathology, support that AKI are different from healthy skin and share features with SCC, implicating that all AKs should be treated because it is not possible to predict their evolution to SCC.
New technologies for the diagnosis and treatment of non-melanoma skin cancers
19-mag-2023
PELLACANI, Giovanni
MAGNONI, Cristina
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11380/1305626
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