Ultra Deep Sequencing of Circulating Cell-Free DNA as a Potential Tool for Hepatocellular Carcinoma Management.

BACKGROUND: Cell-free DNA (cfDNA) concentrations have been described to be inversely correlated with prognosis in cancer. Mutations in HCC-associated driver genes in cfDNA have been reported, but their relation with patient's outcome has not been described. Our aim was to elucidate whether mutations found in cfDNA could be representative from those present in HCC tissue, providing the rationale to use the cfDNA to monitor HCC. METHODS: Tumoral tissue, paired nontumor adjacent tissue and blood samples were collected from 30 HCC patients undergoing curative therapies. Deep sequencing targeting HCC driver genes was performed. RESULTS: Patients with more than 2 ng/µL of cfDNA at diagnosis had higher mortality (mean OS 24.6 vs. 31.87 months, p = 0.01) (AUC = 0.782). Subjects who died during follow-up, had a significantly higher number of mutated genes (p = 0.015) and number of mutations (p = 0.015) on cfDNA. Number of mutated genes (p = 0.001), detected mutations (p = 0.001) in cfDNA and ratio (number of mutations/cfDNA) (p = 0.003) were significantly associated with recurrence. However, patients with a ratio (number of mutations/cfDNA) above 6 (long-rank p = 0.0003) presented a higher risk of recurrence than those with a ratio under 6. Detection of more than four mutations in cfDNA correlated with higher risk of death (long-rank p = 0.042). CONCLUSIONS: In summary, cfDNA and detection of prevalent HCC mutations could have prognostic implications in early-stage HCC patients.

Preclinical In Vivo Validation of the RAD51 Test for Identification of Homologous Recombination-Deficient Tumors and Patient Stratification.

PARP inhibitors (PARPi) are approved drugs for platinum-sensitive, high-grade serous ovarian cancer (HGSOC) and for breast, prostate, and pancreatic cancers (PaC) harboring genetic alterations impairing homologous recombination repair (HRR). Detection of nuclear RAD51 foci in tumor cells is a marker of HRR functionality, and we previously established a test to detect RAD51 nuclear foci. Here, we aimed to validate the RAD51 score cut off and compare the performance of this test to other HRR deficiency (HRD) detection methods. Laboratory models from BRCA1/BRCA2-associated breast cancer, HGSOC, and PaC were developed and evaluated for their response to PARPi and cisplatin. HRD in these models and patient samples was evaluated by DNA sequencing of HRR genes, genomic HRD tests, and RAD51 foci detection. We established patient-derived xenograft models from breast cancer (n = 103), HGSOC (n = 4), and PaC (n = 2) that recapitulated patient HRD status and treatment response. The RAD51 test showed higher accuracy than HRR gene mutations and genomic HRD analysis for predicting PARPi response (95%, 67%, and 71%, respectively). RAD51 detection captured dynamic changes in HRR status upon acquisition of PARPi resistance. The accuracy of the RAD51 test was similar to HRR gene mutations for predicting platinum response. The predefined RAD51 score cut off was validated, and the high predictive value of the RAD51 test in preclinical models was confirmed. These results collectively support pursuing clinical assessment of the RAD51 test in patient samples from randomized trials testing PARPi or platinum-based therapies. SIGNIFICANCE: This work demonstrates the high accuracy of a histopathology-based test based on the detection of RAD51 nuclear foci in predicting response to PARPi and cisplatin.

Liver Histotripsy Mediated Abscopal Effect-Case Report.

We present a case report that shows an abscopal effect in the context of a safety and efficacy clinical trial for histotripsy as ablation technique in liver tumors. The abscopal effect appears in the form of reduction in the volume of nontreated tumor lesions in the same organ, as well as sustained reduction of tumor marker [carcinoembryonic antigen (CEA)] that extends weeks away of the procedure. Histotripsy is a novel noninvasive, nonthermal, and nonionizing precise ablation technique for tissue destruction guided by ultrasonography. We discuss the feasibility of this technique compared with other focal therapies and its possibilities as immune system enhancer.

Contrast-Enhanced Ultrasound: A Useful Tool to Study and Monitor Hepatic Tumors Treated With Histotripsy.

Histotripsy is a novel noninvasive nonthermal, nonionizing, and precise treatment technique for tissue destruction. Contrast-enhanced ultrasound (CEUS) improves the detection, characterization, and follow-up of hepatic lesions because it depicts accurately the vascular perfusion of both normal hepatic tissue and hepatic tumors. We present the spectrum of imaging findings of CEUS after histotripsy treatment of hepatic tumors. CEUS provides real-time information, a close approximation to the dimension of the lesion, and a clear definition of its margins. Hepatic tumors detected by ultrasound can be potentially treated using B-mode ultrasound-guided histotripsy and characterized and monitored with CEUS. CEUS has shown to be very useful after tissue treatment to monitor and assess the evolution of the treated zone. Histotripsy treated zones are practically isoechogenic and slightly heterogeneous, and their limits are difficult to establish using standard B-mode ultrasound. The use of CEUS after histotripsy showing uptake of contrast protruding into the treated zone is clinically relevant to identify residual tumors and establish the most appropriate management strategy avoiding unnecessary treatments. We here describe CEUS findings after histotripsy for hepatic tumors.

Active bleeding after kidney biopsy: Successful ultrasound-guided direct thrombin embolization into the cortical fistula.

A 27-year-old man, previously diagnosed with IgA nephropathy, was referred for native kidney biopsy. After the procedure, the patient presented active bleeding revealed by Doppler and contrast-enhanced ultrasonography at the biopsy site. Successful embolization of the cortical fistula, the focus of bleeding, was achieved using ultrasound-guided thrombin injection and confirmed by Doppler ultrasonography, contrast-enhanced ultrasonography, and CT angiography. This case report shows that contrast-enhanced ultrasonography is useful for detecting active bleeding after a solid organ biopsy. Moreover, ultrasound-guided thrombin embolization is a safe and minimally invasive treatment and an alternative to angiography-guided embolization.

One-step nucleic acid amplification for intraoperative analysis of sentinel lymph node in papillary thyroid carcinoma.

Objective Lymphadenectomy in papillary thyroid carcinoma (PTC) is controversial. It is indicated whenever metastases have been proven before or during surgery and as a prophylactic treatment in high-risk patients. However, 30-50% of cN0 patients become pN1 postoperatively. In PTC, selective-sentinel-lymph-node-biopsy (SLNB) with conventional intraoperative analysis is 8% false negative. One-step nucleic acid amplification (OSNA) is a molecular technique which allows real-time detection of mRNA encoding for cytokeratin 19. OSNA has been introduced in intraoperative analysis of several tumors to reduce false-negative rates and distinguish micrometastasis from macrometastasis. Our objective was to evaluate the impact of the introduction of OSNA in the intraoperative evaluation of the sentinel node (SN) in PTC. Design We analyzed a series of 35 patients subjected to SLNB. Methods All the dissected nodes, SN and non-SN, were evaluated with OSNA and cytology. Results We obtained a total of 110 SN. SLNB proved positive in 14 patients (40%) with cytology and in 23 (65.7%) with OSNA (P < 0.001). In the 29 patients with subsequent lymphadenectomy we obtained 360 lymph nodes ((52 positive in cytology (14.4%) and 107 in OSNA (29.7%)). Lymphadenectomy proved positive in 16 patients according to cytology (55%) and in 24 according to OSNA (83%) (P = 0002). The majority of patients with micrometastasis in SN showed only micrometastasis in lymphadenectomy. Conclusions The present study shows selective-sentinel-lymph-node-biopsy with one-step nucleic acid amplification technique to be feasible in papillary thyroid carcinoma. The quantitative nature of one-step nucleic acid amplification paves the way toward a more personalized surgical approach, limiting lymphadenectomy to patients with intraoperative evidence of macrometastasis in the sentinel node.

A RAD51 assay feasible in routine tumor samples calls PARP inhibitor response beyond BRCA mutation.

Poly(ADP-ribose) polymerase (PARP) inhibitors (PARPi) are effective in cancers with defective homologous recombination DNA repair (HRR), including BRCA1/2-related cancers. A test to identify additional HRR-deficient tumors will help to extend their use in new indications. We evaluated the activity of the PARPi olaparib in patient-derived tumor xenografts (PDXs) from breast cancer (BC) patients and investigated mechanisms of sensitivity through exome sequencing, BRCA1 promoter methylation analysis, and immunostaining of HRR proteins, including RAD51 nuclear foci. In an independent BC PDX panel, the predictive capacity of the RAD51 score and the homologous recombination deficiency (HRD) score were compared. To examine the clinical feasibility of the RAD51 assay, we scored archival breast tumor samples, including PALB2-related hereditary cancers. The RAD51 score was highly discriminative of PARPi sensitivity versus PARPi resistance in BC PDXs and outperformed the genomic test. In clinical samples, all PALB2-related tumors were classified as HRR-deficient by the RAD51 score. The functional biomarker RAD51 enables the identification of PARPi-sensitive BC and broadens the population who may benefit from this therapy beyond BRCA1/2-related cancers.

Paratracheal lymph nodes: a new sonographic finding in autoimmune thyroiditis.

PURPOSE: To assess the clinical value of paratracheal lymph nodes (PLNs) as a novel sonographic finding in autoimmune thyroiditis (AT). METHODS: A total of 309 consecutive patients underwent sonographic examinations of the thyroid between 1998 and 2003. A single radiologist assessed the sonographic findings of AT and PLNs. All patients underwent serological tests for antimicrosomal antibodies (AMAs). Patients with clinical, cytological, or laboratory findings of thyroiditis formed the AT group with a positive AMA test (n = 199). Controls were patients with no signs of nodular thyroid disease, normal thyrotropin, negative AMA, and benign cytology (n = 110). RESULTS: PLNs were seen in 184 of 199 patients in the AT group and in 28 of 110 controls (P < 0.001) (sensitivity of 93.4%, specificity of 74.5% in the diagnosis of AT). PLNs in controls were fewer (2.8 +/- 1.5 versus 4.7 +/- 2.6; P < 0.001) and smaller (8.2 +/- 2.4 mm versus 10.7 +/- 3.3 mm; P < 0.001) than in the AT group. CONCLUSION: PLNs are often present in patients with AT and are detectable with sonography. Radiologists should be aware of the importance of including the paratracheal region in the evaluation of the thyroid gland.

Value of sonography for follow-up of mediastinal lymphadenopathy in children with tuberculosis.

PURPOSE: To assess the clinical value of sonography for the follow-up of mediastinal lymphadenopathy in children diagnosed with pulmonary tuberculosis (TB). METHODS: We conducted a retrospective review of the medical records of 21 children (9 boys, 12 girls) with a mean age of 6 years (range, 7.4 months to 18 years) who had a positive intradermal tuberculin skin test. All patients underwent thorough history-taking, physical examination, frontal and lateral chest radiographs, and sonographic study of the mediastinum. The mediastinum was accessed through the suprasternal and left parasternal approaches. The presence of 1 or more masses with an ovoid or round shape and hypoechoic appearance in the anterior or middle mediastinum was recorded. A comparison was made between the results of the sonographic examination of the mediastinum before administration of anti-TB agents and after 3 months of treatment. RESULTS: Pulmonary radiographic findings were suggestive of TB in 17 patients and were uncertain in 4 patients. Sonographic examination, however, detected mediastinal lymphadenopathy in all patients. A comparison of pretreatment mediastinal sonograms with those obtained after 3 months of anti-TB treatment showed a marked reduction of lymph node involvement in 17 patients (80.9%). In the remaining 4 patients, mediastinal lymphadenopathy was still present. CONCLUSION: Mediastinal sonography appears to be a valuable tool for the diagnosis of TB and in the monitoring of response to treatment in children.

Comparison of ultrasound with plain radiography and CT for the detection of mediastinal lymphadenopathy in children with tuberculosis.

BACKGROUND: Lymphadenopathy, with or without parenchymal abnormality, is the radiological hallmark of primary tuberculosis (TB) in children. However, lymph node enlargement may pass undetected on plain chest radiographs. Ultrasonography provides complementary information to that obtained by radiographs. OBJECTIVE: To assess the clinical value of US for the detection of mediastinal lymphadenopathy in children with a positive intradermal tuberculin test. MATERIALS AND METHODS: Thirty-two children with a mean age of 6 years and a positive Mantoux test underwent chest radiography (frontal and lateral) and US (suprasternal and left parasternal access routes). Chest CT was performed at the discretion of the attending physician in six cases. RESULTS: Eleven children had clinical symptoms and 90% a recent contact with a person with active TB. In 90.5% of children with chest radiographic images compatible with TB, coincident findings in the mediastinal US study were found. By comparison, 66.7% of those with normal chest radiography had evidence of mediastinal lymphadenopathy on the US scan. In all cases but one, US and CT findings agreed. CONCLUSIONS: Mediastinal US is useful for the detection of enlarged lymph nodes in children with a positive tuberculin reaction and normal chest radiography.