Multicancer screening test based on the detection of circulating non haematological proliferating atypical cells.

BACKGROUND: the problem in early diagnosis of sporadic cancer is understanding the individual's risk to develop disease. In response to this need, global scientific research is focusing on developing predictive models based on non-invasive screening tests. A tentative solution to the problem may be a cancer screening blood-based test able to discover those cell requirements triggering subclinical and clinical onset latency, at the stage when the cell disorder, i.e. atypical epithelial hyperplasia, is still in a subclinical stage of proliferative dysregulation. METHODS: a well-established procedure to identify proliferating circulating tumor cells was deployed to measure the cell proliferation of circulating non-haematological cells which may suggest tumor pathology. Moreover, the data collected were processed by a supervised machine learning model to make the prediction. RESULTS: the developed test combining circulating non-haematological cell proliferation data and artificial intelligence shows 98.8% of accuracy, 100% sensitivity, and 95% specificity. CONCLUSION: this proof of concept study demonstrates that integration of innovative non invasive methods and predictive-models can be decisive in assessing the health status of an individual, and achieve cutting-edge results in cancer prevention and management.

Minimal residual disease assessment of papillary thyroid carcinoma through circulating tumor cell-based cytology.

The overall estimated risk of recurrence after an apparently complete thyroid cancer resection ranges from <1% to 55%, and the high-quality pathology report is crucial for proper risk stratification. The neck ultrasound (US) and serum thyroglobulin (Tg) and anti-Tg antibody (TgAb) assays are the mainstays for Differentiated Thyroid Cancer (DTC) follow-up. However, the neck US includes a high frequency of nonspecific findings and despite the serum, Tg unmasks the presence of thyrocytes, it is not discriminating between normal and malignant cells. In this study, to improve post-surgery follow-up of minimal residual disease in papillary thyroid cancer (PTC) patients, blood-derived cytology specimens were evaluated for the presence of circulating tumor cells (CTCs). The presence of CTCs of thyroid origin was confirmed by cytomorphological and tissue-specific antigens analysis (Thyroid Transcription Factor-1/TTF-1 and Tg) and proliferative profile (percentage of cells in S-phase). Our data revealed an unfavorable' prognostic risk in patients with >5% CTCs (p = 0.09) and with >30% S-phase cells at baseline (p = 0.0015), predicting ≤1 year relapsing lesion event. These results suggest a new intriguing frontier of precision oncology forefront cytology-based liquid biopsy.

Tailoring Chemometric Models on Blood-Derived Cultures Secretome to Assess Personalized Cancer Risk Score.

The molecular protonation profiles obtained by means of an organic electrochemical transistor, which is used for analysis of molecular products released by blood-derived cultures, contain a large amount of information The transistor is based on the conductive polymer PEDOT:PSS comprising super hydrophobic SU8 pillars positioned on the substrate to form a non-periodic square lattice to measure the state of protonation on secretomes derived from liquid biopsies. In the extracellular space of cultured cells, the number of glycation products increase, driven both by a glycolysis metabolism and by a compromised function of the glutathione redox system. Glycation products are a consequence of the interaction of the reactive aldehydes and side glycolytic products with other molecules. As a result, the amount of the glycation products reflects the anti-oxidative cellular reserves, counteracting the reactive aldehyde production of which both the secretome protonation profile and cancer risk are related. The protonation profiles can be profitably exploited through the use of mathematical techniques and multivariate statistics. This study provides a novel chemometric approach for molecular analysis of protonation and discusses the possibility of constructing a predictive cancer risk model based on the exploration of data collected by conventional analysis techniques and novel nanotechnological devices.

BRCA1 5083del19 mutant allele selectively up-regulates periostin expression in vitro and in vivo.

PURPOSE: The aim of this study was to explore the gene expression pattern produced by the cancer-associated BRCA1 5083del19 founder mutation by using a microarray analysis. Such a mutation, identified in a subset of familial breast cancer patients, involves a deletion at the 3' end of the BRCA1 messenger leading, in the mature protein, to the ablation of the BRCT tandem domain. EXPERIMENTAL DESIGN: We generated HeLa cells stably expressing both exogenous wild-type (HeLa/(wt)BRCA1), used as a control, and 5083del19 BRCA1 (HeLa/(5083del19)BRCA1) alleles; gene chips were then used to investigate any changes in the transcription profile induced by the 5083del19 BRCA1 mutant compared with controls. RESULTS: Among the genes showing perturbation of their expression, periostin was found to be up-regulated in HeLa/(5083del19)BRCA1 cells to an extent of 72-fold versus HeLa/(pcDNA3.1/empty) and 76-fold versus HeLa/(wt)BRCA1 cells. This finding was validated both in vitro in breast cancer cell lines harboring mutations of BRCA1 and in vivo by immunohistochemistry of breast cancer specimens bearing the 5083del19 BRCA1 mutation as well as by Western blot analysis of sera obtained from patients and healthy carriers of the same mutation. CONCLUSIONS: Our results suggest that periostin overexpression, whose product is released from cells in the extracellular fluids, might be a potential marker for early cancer detection in a specific subset of hereditary breast carcinomas triggered by cancer-associated BRCA1 mutations that affect the BRCT tandem domain.

A novel missense germline mutation in exon 2 of the hMSH2 gene in a HNPCC family from Southern Italy.

Germline mutations within the mismatch repair (MMR) genes are generally found in colorectal cancer (CRC) patients with a positive family history for the presence of the neoplasia. Clinical standard criteria have been established to define hereditary-non-polyposis-colorectal cancer (HNPCC)-prone families. Interestingly, the number of MMR gene mutations found in kindreds not fulfilling these criteria is still increasing. In this work we report the identification of a novel germline mutation of the hMSH2 gene, in two CRC-bearing subjects. The two probands belong to a large kindred from South Italy with no history suggestive for cancer aggregation. On the other hand, the early-onset of the neoplasia as well as the presence of a high number of tumor infiltrating lymphocytes (TILs) in the histological specimens of both patients, prompted us to perform a comprehensive genetic analysis. This analysis included the evaluation of the microsatellite instability (MSI) status with five markers according to the National Cancer Institute recommendations, followed by direct sequencing of the hMLH1 and hMSH2 genes. Both probands were found to carry a germline missense (277 C>T) mutation leading to the change (L93F) of an amino acid residue in a highly conserved domain of the MSH2 protein. This mutation is accompanied by the loss of expression of the hMSH2 gene in the tumor tissue. Our findings suggest that in the presence of the above-mentioned criteria it may be useful to perform a molecular analysis of the MMR genes, even if the pedigree does not show marked aggregation of cancers.