Transcriptome analyses allow the distinguishing of pancreatic ductal adenocarcinoma (PDAC) subtypes, exhibiting different prognoses and chemotherapy responses. However, RNA extraction from pancreatic tissue is cumbersome and has been performed mainly from surgical samples, which are representative of < 20% of cases. The majority of PDAC patients undergo endoscopic ultrasound (EUS)-guided tissue acquisition (EUS-TA), but RNA has been rarely extracted from EUS-TA with scanty results. Herein, we aimed to determine the best conditions for RNA extraction and analysis from PDAC EUS-TA samples in order to carry out molecular analyses. PDAC cases underwent diagnostic EUS-TA, with needles being a 25G fine needle aspiration (FNA) in all patients and then either a 20G lateral core-trap fine needle biopsy (FNB) or a 25G Franseen FNB; the conservation methods were either snap freezing, RNALater or Trizol. RNA concentration and quality (RNA integrity index; RIN) were analyzed and a panel of genes was investigated for tissue contamination and markers of molecular subtype and aggressivity through qRT-PCR. Seventy-four samples from 37 patients were collected. The median RNA concentration was significantly higher in Trizol samples (10.33 ng/uL) compared with snap frozen (0.64 ng/uL; p < 0.0001) and RNALater (0.19 ng/uL; p < 0.0001). The RIN was similar between Trizol (5.15) and snap frozen samples (5.85), while for both methods it was higher compared with RNALater (2.7). Among the needles, no substantial difference was seen in terms of RNA concentration and quality. qRT-PCR analyses revealed that samples from all needles were suitable for the detection of PDAC subtype markers (GATA6 and ZEB1) and splice variants associated with mutational status (GAP17) as well as for the detection of contaminating tissue around PDAC cells. This is the first study that specifically investigates the best methodology for RNA extraction from EUS-TA. A higher amount of good quality RNA is obtainable with conservation in Trizol with a clear superiority of neither FNA nor FNB needles. RNA samples from EUS-TA are suitable for transcriptome analysis including the investigation of molecular subtype and splice variants expression.

Lung cancer is one of the most common solid malignant disease and an accurate staging is required for planning the most appropriate treatment. Surgery or local radiotherapy are the elective treatment for localized disease, however, the presence of mediastinal malignant lymph nodes can affect the role of surgery as a first-line therapy and a chemotherapy and/or radiotherapy treatment are recommended. Currently endoscopic ultrasound (EUS) in the mediastinum has brought the important advantage of a minimal-invasive diagnostic approach for the diagnosis and staging of malignant conditions, having the possibility of sampling lesions or lymph nodes in real-time. EUS’ diagnostic capabilities have increased over time and currently it represents an excellent approach for the staging of lung cancer, especially combined to endobronchial ultrasound (EBUS). Last generation EUS equipments (echoendoscopes, ultrasound processors, needles) and tools [contrast-enhanced EUS (CH-EUS), elastography] can help the endosonographer during the ultrasound evaluation and they can increase the accuracy of lung cancer staging. A systematic method of evaluation of all the visible lymph nodes stations is fundamental to complete the lung cancer staging, as well as a correct endoscopic room organization of the equipments, healthcare personnel and devices.

In recent years, small pancreatic neuroendocrine tumors (pNETs) have shown a dramatic increase in terms of incidence and prevalence, and endoscopic ultrasound (EUS) radiofrequency ablation (RFA) is one potential method to treat the disease in selected patients. As well as the heterogeneity of pNET histology, the studies reported in the literature on EUS-RFA procedures for pNETs are heterogeneous in terms of ablation settings (particularly ablation powers), radiological controls, and radiological indications. The aim of this review is to report the current reported experience in EUS-RFA of small pNETs to help formulate the procedure indications and ablation settings. Another aim is to evaluate the timing and the modality of the radiological surveillance after the ablation. Moreover, new studies on large-scale series are needed in terms of the safety and long-term oncological efficacy of RFA on these small lesions.

(1) Background: Preclinical and clinical data about a novel radiofrequency ablation (RFA) system (STARmed Co, Ltd.; Koyang, Korea) designed to be used under endoscopic ultrasound (EUS) control for pancreatic lesion ablation, are limited, obtained with non-standardized procedures and heterogeneous results. The aim of this study is to standardize the RFA procedure of this system in order to define the optimal ablation power and time. (2) Methods: RFA was performed on an ex-vivo porcine liver at different powers (40, 30, 20, 10 Watts (W)) and times (1, 3, 5, 7, 15 min) with a 1-centimeter monopolar electrode (perfused by chilled solution) positioned on the distal tip of a 19-Gauge needle. A blinded expert pathologist histologically analyzed each ablation area. (3) Results: The size of the total macroscopic ablated area was negatively correlated with ablation power (R −0.74): the largest was obtained at 10 W (p = 4.7 × 10⁻⁴) for longer times (R 0.92; p = 8.9 × 10⁻⁸). Central histologic coagulative necrosis did not differ among ablation settings (mean size 3.25 mm). External “parenchymal hypochromia” or “diaphanization” resulted the widest at 10 W, for longer times (R 0.8, p = 3.6 × 10⁻⁴). (4) Conclusions: The RFA system can produce small sizes of coagulative necrosis, regardless of the setting. Larger areas of diaphanization surrounding the necrosis can be produced at lower powers for longer times.

Endoscopic ultrasound-ablation with HybridTherm-Probe (EUS-HTP) significantly reduces tumour volume (TV) in locally-advanced pancreatic ductal adenocarcinoma (LA-PDAC). We aimed at investigating the clinical efficacy of EUS-HTP plus chemotherapy versus chemotherapy (HTP-CT and CT arms) in LA- and borderline-resectable (BR) PDAC, with 6-months progression-free survival (6-PFS) rate as primary endpoint. In a phase-II randomized-controlled-trial, 33 LA/BR-PDAC patients per-arm were planned to verify 20% improved 6-PFS rate. Radiological response (Choi criteria), TV and serum CA19.9 were assessed up to 6-months. Seventeen and 20 LA/BR-PDAC patients were randomized to HTP-CT or CT. Baseline and CT-related features were balanced. At 6-months, 6-PFS rate was 41.2% and 30% in HTP-CT and CT arms (p = 0.48), respectively. A decrease ≥50% of serum CA19.9 was achieved in 75% and 64.3% of HTP-CT and CT patients (p = 0.53), respectively. TV reduced up to 6-months in 64.3% and 47.1% of HTP-CT and CT patients (p = 0.35), respectively. Resection rate, PFS-time and overall survival (OS-time) were similar. HTP-CT achieves a non-significant 11.2%, 10.7% and 17.2% improved 6-PFS, CA19.9 decrease ≥50% and TV reduction rates over CT, without any impact on resection rate, PFS-time and OS-time. As the study was underpowered, these results suggest further investigation of EUS-local ablation in selected patients with localized disease after induction CT.

Gastric cancer in 2022: Is there still a role for endoscopic ultrasound?