Backgroud: Neoplastic and preneoplastic lesions of the uterine cervix represent a significant social and health problem, still too many women come to the observation with advanced lesions that require invasive treatments. The morphological analysis has low sensitivity and variability. Only integration with molecular data would allow greater diagnostic robustness, a personalization of drug treatment and follow-up. Purpose: The goal of this study is to improve the characterization of lesions at the second level through non-morphological analyzes. Persistent infection is necessary, but not sufficient to induce neoplastic transformation. The presence of cofactors capable of influencing the progress of the lesion is required, such as: the genotype, the viral load, the presence of coinfections and the integration status of the HPV genome in host cell DNA. However, first-level screening tests alone are unable to determine them as they are based on morphology. The use of a molecular test to detect cofactors in order to improve diagnostics at the second level, improve the characterization of the lesion and understand the real risk of progression appears necessary. This would allow targeted treatment of only women with lesions at high risk of progression. Materials and methods: The analysis of cervical samples is performed on FFPE material, especially biopsies and courettage. The samples were analyzed in duplicate: both in immunohistochemistry to evaluate the expression of p16 and Ki67; and from the molecular point of view through a multiplex Real time platform for the determination of viral load, genotype and coinfections. Results: From the histological analysis it emerged that the 55 samples analyzed are 22/55 CIN1 cases and of these the molecular analysis highlights 12/22 negative cases; so, 54% of CIN1 actually are not HPV-related lesions. Furthermore, 10 cases of CIN1 are co-infected and with high viral loads and are at greater risk of neoplastic progression and should be closely monitored over time. Discussion: The study confirms that the morphological analysis alone tends to overestimate the CIN1, with repercussions above all for the patients, who undergo further diagnostic tests, including invasive ones, when, on the other hand, they would not be treated. Furthermore, it was possible to detect low-grade lesions characterized by co-infections, with high-risk, high-burden strains, making them at high risk of progression. These cases should be monitored over time. Conclusion: Molecular analysis can improve diagnosis and allows for a personalized therapeutic protocol. Furthermore, it is necessary to refine the diagnostic procedures in the second level in order to send only the patients who really need it to conization The experience gained will be extended to other anatomical areas affected by HPV-related pathology.
Backgroud: Le lesioni neoplastiche e preneoplastiche della cervice uterina rappresentano un rilevante problema sociosanitario, ancora troppe donne giungono all’osservazione con lesioni avanzate che richiedono trattamenti invasivi. L’analisi morfologica ha scarsa sensibilità e variabilità. Solo l’integrazione con dati molecolari consentirebbe una maggiore robustezza diagnostica, una personalizzazione del trattamento farmacologico e di follow up. Scopo: L’obiettivo di questo studio è quello di migliorare la caratterizzazione delle lesioni al secondo livello attraverso analisi non morfologiche. L’infezione persistente è necessaria, ma non sufficiente per indurre la trasformazione neoplastica. Occorre la presenza di cofattori capaci di influenzare l’andamento della lesione, come: il genotipo, la carica virale, la presenza di coinfezioni e lo status di integrazione del genoma di HPV in quello della cellula ospite. Tuttavia, i soli esami di screening di primo livello non sono in grado di determinarli in quanto basati sulla morfologia. Si propone l’uso di un test molecolare in grado di rilevare i cofattori in modo da migliorare la diagnostica al secondo livello, caratterizzare la lesione capendone il reale rischio di progressione. Ciò consentirebbe di trattare in modo mirato solo le donne con lesioni ad alto rischio di progressione. Materiali e metodi: L’analisi dei campioni cervicali è eseguita su materiale FFPE, soprattutto di biopsie e courettage. I campioni sono stati analizzati in doppio: sia in immunoistochimica per valutare l’espressione di p16 e Ki67; sia dal punto di vista molecolare tramite piattaforma multiplex Real time per la determinazione di carica virale, genotipo e coinfezioni Risultati: Dall’analisi istologica è emerso che i 55 campioni analizzati sono 22/55 casi CIN1 e di questi l’analisi molecolare evidenzia 12/22 casi negativi, ovvero il 54% dei CIN1 in realtà non sono lesioni HPV-relate. Inoltre, 10 casi di CIN1 sono coinfettati e con cariche virali alte, ovvero sono a maggior rischio di progressione neoplastica e andrebbero strettamente monitorati nel tempo. Discussione: Lo studio conferma che con la sola analisi morfologica si tende a sovrastimare le CIN1, con ripercussioni soprattutto per le pazienti, che vanno incontro ad ulteriori accertamenti diagnostici, anche di tipo invasivo, quando, invece, non sarebbero da trattare. Inoltre, è stato possibile rilevare lesioni di basso grado caratterizzate da coinfezioni, con ceppi ad alto rischio e carica alta, rendendole ad alto rischio di progressione. Questi casi andrebbero monitorati nel tempo. Conclusione: L’analisi molecolare può migliorare la diagnosi e consente un protocollo terapeutico personalizzato. Inoltre, occorre affinare le procedure diagnostiche nel secondo livello in modo da inviare a conizzazione solo le pazienti che ne hanno reale bisogno L’esperienza acquisita verrà estesa ad altri distretti anatomici colpiti da patologia HPV-relata.
Tipizzazione morfo-molecolare nelle lesioni preneoplastiche della cervice uterina
VARESANO, SERENA
2022-04-12
Abstract
Backgroud: Neoplastic and preneoplastic lesions of the uterine cervix represent a significant social and health problem, still too many women come to the observation with advanced lesions that require invasive treatments. The morphological analysis has low sensitivity and variability. Only integration with molecular data would allow greater diagnostic robustness, a personalization of drug treatment and follow-up. Purpose: The goal of this study is to improve the characterization of lesions at the second level through non-morphological analyzes. Persistent infection is necessary, but not sufficient to induce neoplastic transformation. The presence of cofactors capable of influencing the progress of the lesion is required, such as: the genotype, the viral load, the presence of coinfections and the integration status of the HPV genome in host cell DNA. However, first-level screening tests alone are unable to determine them as they are based on morphology. The use of a molecular test to detect cofactors in order to improve diagnostics at the second level, improve the characterization of the lesion and understand the real risk of progression appears necessary. This would allow targeted treatment of only women with lesions at high risk of progression. Materials and methods: The analysis of cervical samples is performed on FFPE material, especially biopsies and courettage. The samples were analyzed in duplicate: both in immunohistochemistry to evaluate the expression of p16 and Ki67; and from the molecular point of view through a multiplex Real time platform for the determination of viral load, genotype and coinfections. Results: From the histological analysis it emerged that the 55 samples analyzed are 22/55 CIN1 cases and of these the molecular analysis highlights 12/22 negative cases; so, 54% of CIN1 actually are not HPV-related lesions. Furthermore, 10 cases of CIN1 are co-infected and with high viral loads and are at greater risk of neoplastic progression and should be closely monitored over time. Discussion: The study confirms that the morphological analysis alone tends to overestimate the CIN1, with repercussions above all for the patients, who undergo further diagnostic tests, including invasive ones, when, on the other hand, they would not be treated. Furthermore, it was possible to detect low-grade lesions characterized by co-infections, with high-risk, high-burden strains, making them at high risk of progression. These cases should be monitored over time. Conclusion: Molecular analysis can improve diagnosis and allows for a personalized therapeutic protocol. Furthermore, it is necessary to refine the diagnostic procedures in the second level in order to send only the patients who really need it to conization The experience gained will be extended to other anatomical areas affected by HPV-related pathology.File | Dimensione | Formato | |
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