Microfluidic organ-on-chip system for multi-analyte monitoring of metabolites in 3D cell cultures

An organ-on-chip platform equipped with microsensors for long-term microfluidic cultivation and metabolic monitoring (O2, Glu, Lac) of 3D tumour organoid cultures grown from patient-derived single cancer stem cells.

Recurrent Superenhancer of the Oncogene POU5F1B in Colorectal Cancers

Superenhancer usages in single cancer form such as colorectal cancer (CRC) may provide novel efficient targeting candidates. It is unclear whether CRC contains recurrent superenhancers that confer a predisposition to malignancy. We investigated the superenhancer profile of CRC cell line HCT116 and compared it to that of a healthy sigmoid colon. We found that HCT116 had lost most of the normal colon superenhancer activities but gained a new set of tumor-favoring superenhancers that facilitate tumor proliferation, growth signalling, and hypoxia resistance. Inhibiting the superenhancers by JQ-1 treatment had significantly decreased the colony formation capability of HCT116. Then, by comparing the superenhancer genes and robust CRC upregulated genes, we identified a superenhancer associated with a common CRC upregulated oncogene, POU5f1B. POU5f1B overexpression is related to the worse outcome in CRCs. Via performing ChIP-PCR in 35 clinical samples and investigating CRC anti-H3K27ac ChiP-seq public dataset consisting of 36 samples, we further identified that the superenhancer of oncogene POU5F1B is recurrently activated in CRCs, taking 62 and 72 per cent, respectively. Moreover, JQ-1 treatment successfully inhibited the POU5F1B expression in 5 out of 6 POU5F1B superenhancer-positive samples. Therefore, we concluded that the superenhancer activation of POU5F1B contributes partially to its high expression in CRCs, in addition to the well-known gene amplification aetiology.

Measurement of the Drug Sensitivity of Single Prostate Cancer Cells

The treatment of cancer faces a serious challenge as cancer cells within patients are heterogeneous and frequently resistant to therapeutic drugs. Here, we introduce a technology enabling the assessment of single cancer cells exposed to different drugs. PCa cells were individually sorted in self-seeding microwells, cultured for 24 h, and then exposed to several drugs to induce (R1881) or inhibit (Enzalutamide/Abiraterone) the secretion of a protein (PSA). Cell viability and PSA secretion of each individual prostate cell were monitored over a 3-day period. The PSA protein secreted by each cell was captured on a PVDF membrane through a pore in the bottom of each well. The basal PSA secretion was found to be 6.1 ± 4.5 and 3.7 ± 1.9 pg/cell/day for LNCaP and VCaP, respectively. After exposure to R1881, the PSA secretion increased by ~90% on average and was not altered for ~10% of the cells. PSA production decreased in the majority of cells after exposure to enzalutamide and abiraterone.

Follicular Lymphoma in a Single Cancer Center and Validation of FLIPI, FLIPI2, PRIMA and POD24 in Non-Caucasian Patients from Peru

Background Follicular lymphoma (FL) is the second most common non-Hodgkin lymphoma subtype seen in Caucasian countries. Recent data were published on FL from Latin America (LATAM), presenting information of 12 countries. However, this information is widely variable because of the diversity of the Latin population (Caucasian and mestizo individuals). Our aim was to evaluate the clinical features, treatment patterns outcomes of non-Caucasian patients with FL from a single cancer center. Our second aim was to validate FLIPI1, FLIPI2, PRIMA and POD-24 score in our cohort. Methods: This is a retrospective study, including all patients with a pathological diagnosis of FL at the National Institute of Neoplastic Diseases in Lima, Peru from 2010 to 2019. All cases were reviewed by specialized pathologists. Baseline clinical and pathological data were collected. Responses were assessed based on the Lugano criteria. Overall survival (OS) was estimated using the Kaplan-Meier method. Differences were compared with the log-rank test. Results From 2010 to 2019 4480 patients with B-cell lymphoma were diagnosed, 449 patients (10%) had grade 1 to 3A FL, 302 patients had the five variables for FLIPI, 242 for FLIPI2, 257 for the 2 variables for PRIMA prognostic index and 209 received systemic treatment and had enough information to evaluate POD24. The median age for the entire cohort was 59 years old (range 24-92), 49% of patients were ≥60 years, 46% were male, 22% had extranodal involvement, 34% had bulky disease (≥6 cm in diameter), 68% had stage III/IV disease, 32% had hemoglobin <12 g/dl, 8% had serum albumin <3 g/dl, 35% had elevated serum LDH, 33% had B2-microglobulin ≥3,5 mg/l, 22% had bone marrow involvement and 23% had lymph node sites >4. Low, intermediate and high FLIPI were seen in 39%, 27% and 34% of patients, respectively. Low, intermediate and high-risk FLIPI2 was seen in 23%, 54% and 23% of patients, respectively. Low, intermediate, and high PRIMA was seen in 53%, 12% and 35%, respectively. 209 patients received systemic treatment, 60% received CHOP ± rituximab (R), 13% CVP ± R, 15% CHOP, 11% CVP. Response data were available in 158 patients with complete response in 35%, partial response in 57% and no response in 8%, for an overall response rate of 92%. 19% of patient had disease progression within 24 months of first treatment initiation (POD24). For the entire cohort, the median follow-up time was 2.6 years (Interquartile range [IQR] 0.08-13.6), the median OS was not reached (IQR 4.2-not reached). 5y was OS 72% (95% CI 65-77). 5y OS for low, intermediate and high FLIPI were 90% (95% CI 81-94.5), 65% (95% CI 47.3-78.2) and 60.1% (95% CI 46.6-72.4), respectively (p<0.001; Figure a). 5y OS for low, intermediate and high FLIPI2 were 91.9% (95% CI 76.3-97.3), 75.4% (95% CI 63.8-83.7) and 52.9% (95% CI 35.5-67.6), respectively (p<0.001; Figure b). 5y OS for low, intermediate and high PRIMA were 80.5% (95% CI 70-87.6), 79.4% (95% CI 52-92), and 61% (95% CI 47-73), respectively (p=0.004; Figure c). Patients who had and did not have POD24 had median OS of 5.7 years (IQR 2-NR) and NR (IQR 5.1-NR), respectively (p<0.001). 5y OS for patients who had and did not have POD24 was 54.1 % (95% CI 31-63) and 75.2% (95% CI 66-82), respectively (p=0.01). Conclusion: FL has a lower incidence in non-Caucasian patients in Peru compared to those reported for Western countries or other LATAM Caucasian population. FL patients showed higher rates of high FLIPI and FLIPI2 than previously reported in the literature. Chemoimmunotherapy is the standard approach to FL patients, which is associated with high rates of overall response, but low rate of complete response. The OS rates are shorter than those reported in the previously LATAM cohort. Our study validates the prognostic value of FLIPI1, FLIPI2, PRIMA and POD24. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

Unexpected High Frequency of Pathogenic Germline Variants in Older Adults with Primary Myelodysplastic Syndrome

Background: Germline predisposition is increasingly being recognised in myeloid neoplasms (MN) including primary myelodysplastic syndrome. An unequivocal diagnosis of germline predisposition carries actionable considerations for patient management including donor stem cell source for allogeneic transplantation, dose-reduction of conditioning regimes and screening for extra-hematological disease (such as pulmonary abnormalities in patients with telomere biology disorders). In addition, the identification of MDS predisposition syndromes can avoid misdiagnosis (for example, distinguishing idiopathic thrombocytopenic purpura from thrombocytopenia due to RUNX1 germline variant). However, the prevalence of pathogenic germline variants (PGVs) in unselected pMDS patients presenting at older age remains unknown. Aim: This study assesses frequency and type of pathogenic germline variants in MDS patients and compares with age matched healthy controls and patients with other cancers. Method: We analysed 68 known cancer predisposition genes in germline samples of 146 samples from myeloid neoplasms. Study included primary MDS (n=51) and MDS diagnosed in cancer survivors with (n=77) or without prior exposure to cytotoxic therapy (n=18). Using uniform American College of Medical Genetics and Genomics (ACMG) guidelines for annotating germline mutation, we also compared the frequency of pathogenic germline variants in the same genes with patients with single cancer and age-match healthy controls (>70 years). Results: Pathogenic germline variants (PGVs) were identified in 19% (28/146) patients compared to 4% and 3% patients with single cancer and age-matched controls respectively (P<0.0001) (Fig. 1A). Median age at diagnosis was similar between MN patients with or without PGVs [66 years (19-81) vs. 70 years (33-87); P=0.06]. PGVs were most frequent in DDX41 (n=7, 33%) followed by BRCA1 (n=2, 10%), GATA2 (n=2; 10%) and TP53 (n=2; 10%) (Fig.1B). We also identified pathogenic copy number variations (CNV) in 4 patients. The distribution of PGVs was also different, with DDX41 PGVs absent in single cancers and more prevalent in MN than age-matched controls (35% vs. 4%, P<0.001). The frequency of PGV was not significantly different between P-MN and T-MN/ MC-MN (17% versus 10%, P = 0.32 (Fig.1C). The frequency of PGV was 30%, 6%, 19%, 15% and 18% in patients ≤50, 51-59, 60-69, 70-79 and >80 years of age (Fig. 1D). Phenotypic features such as monocytopenia and mycobacterium infections (MonoMAC; SA460) and personal and family history of pulmonary fibrosis (SA918) were present in only two cases with PGVs. Family history of MDS/AML was present in only in four cases with PGVs, in which PGVs were found in typical myeloid malignancy genes (DDX41, GATA2). Importantly, some patients with family history of solid cancers carried PGVs in genes traditionally associated with solid cancers (e.g. MSH6, NF1, TP53 and BRCA1). SA927 had a PGV in MSH6 and multiple first-degree relatives with solid cancers including colon, renal and brain cancers. Moreover, 41% of adults with hematological disorders and a personal and/or family history of interstitial lung disease had PGVs in telomere biology disorder genes. Hence, family history should not be restricted to hematological disorders, but also solid cancers and non-malignant phenotypes (e.g. hepatic and pulmonary fibrosis). The frequency of PGVs was not different in patients with and without family history of cancer (23% vs. 13%, P=0.32). Conclusion: The frequency of PGVs is significantly high in MN compared to age matched healthy control and other cancer patients. Our observation of a high frequency of PGVs in the older MDS population warrants standardization of germline testing at diagnosis to guide optimal management of patients and their families. Figure 1 Figure 1. Disclosures Hiwase: Novartis: Membership on an entity's Board of Directors or advisory committees; AbbVie: Membership on an entity's Board of Directors or advisory committees.

Combination of terbium-161 with somatostatin receptor antagonists—a potential paradigm shift for the treatment of neuroendocrine neoplasms

Purpose The β¯-emitting terbium-161 also emits conversion and Auger electrons, which are believed to be effective in killing single cancer cells. Terbium-161 was applied with somatostatin receptor (SSTR) agonists that localize in the cytoplasm (DOTATOC) and cellular nucleus (DOTATOC-NLS) or with a SSTR antagonist that localizes at the cell membrane (DOTA-LM3). The aim was to identify the most favorable peptide/terbium-161 combination for the treatment of neuroendocrine neoplasms (NENs). Methods The capability of the 161Tb- and 177Lu-labeled somatostatin (SST) analogues to reduce viability and survival of SSTR-positive AR42J tumor cells was investigated in vitro. The radiopeptides’ tissue distribution profiles were assessed in tumor-bearing mice. The efficacy of terbium-161 compared to lutetium-177 was investigated in therapy studies in mice using DOTATOC or DOTA-LM3, respectively. Results In vitro, [161Tb]Tb-DOTA-LM3 was 102-fold more potent than [177Lu]Lu-DOTA-LM3; however, 161Tb-labeled DOTATOC and DOTATOC-NLS were only 4- to fivefold more effective inhibiting tumor cell viability than their 177Lu-labeled counterparts. This result was confirmed in vivo and demonstrated that [161Tb]Tb-DOTA-LM3 was significantly more effective in delaying tumor growth than [177Lu]Lu-DOTA-LM3, thereby, prolonging survival of the mice. A therapeutic advantage of terbium-161 over lutetium-177 was also manifest when applied with DOTATOC. Since the nuclear localizing sequence (NLS) compromised the in vivo tissue distribution of DOTATOC-NLS, it was not used for therapy. Conclusion The use of membrane-localizing DOTA-LM3 was beneficial and profited from the short-ranged electrons emitted by terbium-161. Based on these preclinical data, [161Tb]Tb-DOTA-LM3 may outperform the clinically employed [177Lu]Lu-DOTATOC for the treatment of patients with NENs.

696 LONG-TERM OUTCOMES OF MINIMALLY INVASIVE ESOPHAGECTOMY COMPARED TO OPEN ESOPHAGECTOMY: A RETROSPECTIVE ANALYSIS OF 2958 CASES IN SINGLE CANCER CENTER

Esophageal cancer is the fourth primary cause of cancer-related death in the male in China.The cornerstone of treatment for resectable esophageal cancer is surgery. With the development of minimally invasive esophagectomy (MIE), it is gradually adopted as an alternative to open esophagectomy (OE) in real-world practice. The purpose of this study is to explore whether MIE vs. OE will bring survival benefits to patients with the advancement of treatment techniques and concepts. Methods Data were obtained from the Sichuan Cancer Hospital & Institute Esophageal Cancer Case Management Database (SCH-ECCM Database). We retrospective analyzed esophageal cancer patients who underwent esophagectomy from Jan. 2010 to Nov. 2017. Patients were divided into two groups: MIE and OE groups. Clinical outcome and survival data were compared using TNM stages of AJCC 8th edition. Results After 65.3 months of median follow-up time, 2958 patients who received esophagectomy were included. 1106 of 2958 patients (37.4%) were underwent MIE, 1533 of 2958 patients (51.8%) were underwent OE. More than half of the patients (56.7%, 1673/2958) were above stage III. The median overall survival (OS) of 2958 patients was 51.6 months (95% CI 45.2–58.1). The MIE group's median OS was 74.6 months compared to 42.4 months in the OE group (95% CI 1.23–1.54, P < 0.001). The OS at 1, 3, and 5 years were 90%, 68%, 58% in the MIE group; 85%, 54%, 42% in the OE group,respectively (P<0.001). Conclusion The nearly 8-year follow-up data from this single cancer center suggests that with the advancement of minimally invasive surgical technology, MIE can bring significant benefits to patients' long-term survival compared with OE. Following the continuous progression of minimally invasive surgery and establishing a mature surgical team, MIE should be encouraged.