Cell Proliferation In Patients Research Articles

METTL3 facilitates stemness properties and tumorigenicity of cancer stem cells in hepatocellular carcinoma through the SOCS3/JAK2/STAT3 signaling pathway.

Liver cancer stem cells (LCSCs) contribute to tumor recurrence and cancer cell proliferation in patients with hepatocellular carcinoma (HCC). METTL3-catalyzed m6A modification is relevant to the cancer stem cell (CSC) phenotype, including LCSCs. LCSCs were isolated from MHCC-97H and HepG2 cells through flow cytometry. UALCAN data were used to analyze the expression of METTL3 in liver hepatocellular carcinoma (LIHC) tissues. Loss- and gain-of-function experiments were utilized to assess the biological effects of METTL3 and SOCS3 on the proliferation and stemness phenotypes in vitro and in vivo. The mechanisms underlying the impact of METTL3 were explored using qPCR, MeRIP-qPCR, dual-luciferase reporter, and western blot assays. METTL3 was significantly upregulated in LIHC tissues according to the UALCAN database. METTL3 was highly expressed in LIHC and was significantly correlated with individual cancer stage, tumor grade and lymph node metastasis. Patients with low METTL3 expression had a longer overall survival time based on the data from UALCAN. In addition, the level of METTL3 was enhanced in LCSCs and decreased in non-LCSCs compared to HCC cells. Moreover, overexpression of METTL3 stimulated the proliferation and stemness of LCSCs in vitro and in vivo, while loss of METTL3 impeded it. Bioinformatics analysis combined with validation experiments determined that m6A was modified by METTL3-targeting SOCS3 mRNA. METTL3 had side effects regarding the stability of SOCS3 mRNA. SOCS3 overexpression impaired and SOCS3 depletion facilitated the development of LCSCs via the JAK2/STAT3 pathway. Furthermore, METTL3 depletion suppressed proliferation and stemness in LCSCs, which was restored by SOCS3 knockdown or colivelin treatment. We discovered that METTL3 facilitated the stemness and tumorigenicity of LCSCs by modifying SOCS3 mRNA with m6A.

O-119 - Cutaneous manifestation of mature plasmacytoid dendritic cell proliferation in patients with myeloid leukemia

O-119 - Cutaneous manifestation of mature plasmacytoid dendritic cell proliferation in patients with myeloid leukemia

Abstract P3-03-27: Assessment of the proliferative activity of breast tissue after the administration of combined oral hormonal contraceptive, associated or not with estriol

Abstract The molecular mechanisms related to the breast cancer development are poorly understood, particularly those associated with estrogen. Evidence is mounting that developmental differences may influence the risk of cancer. However, understanding breast development, morphology and the biochemical factors that influence them is pertinent to study pre-malignant and malignant conditions that affect the breasts. Objective: To evaluate the proliferative activity of normal breast epithelium, through the c-myc expression, after the administration of combined oral hormonal contraceptive, associated or not with estriol. Retrospective double-blind study, in which the effect of combined oral contraceptives on the normal breast epithelium in women with fibroadenoma was evaluated. We analyzed 33 women In a cohort of 70 selected (37 excluded). The control group (1) received a compound containing levonorgestrel 0.15mg (LNG) and ethinylestradiol 0.03mg (EE), associated with 2mg of placebo (PLC) manufactured in the same capsule ingested daily for 21 days with an interval of 7 days, during four cycles. The study group (2) received a compound containing levonorgestrel 0.15 mg (LNG) and ethinylestradiol 0.03 mg (EE), associated with estriol 2 mg (E3) manufactured in the same capsule ingested daily for 21 days with an interval of 7 days, up to four cycles, when was proceeded the lumpectomy plus normal breast tissue sample. The UNIFESP-EPM Department of Pathology received the samples collected and fixed in 10% buffered formalin. We used an automation device to perform the Immunohistochemistry reactions, using c-myc antibodies, added DAB developer (diaminobenzidine) that provides a bright brown color that provides good contrast. We used conventional optical microscopy to read TMA slides. We considered positive the nuclei stained with dark brown in contrast to the blue negative to the c-myc reactions. We evaluated the epithelial areas at least five acinar units were present in the sample at 40X magnification. In the statistical analysis, the initial stages of consolidation as well as the analysis were performed in Software R (2020) and Rstudio (version 4.1), using the packages Tidyverse (2016), Psyco, SummaryTools, Janitor and DataExplorer. After verifying the assumptions of normality, independence and linearity, Student’s T test was performed for independent samples. An alpha of 0.05 was chosen for rejection of the null hypothesis, that is, a confidence interval of the test results of 95%. An effect size measure of the difference between the groups was also performed, called Cohen’s D; it ranges from -1 to 1, and the closer to 0, the smaller the effect size of the difference between the groups. Regarding the control group (30.30%) and the case group (69.70%). The table 2 presents the results for group 1 (COC + Placebo), while table 3 presents the results for group 2 (COC + Estriol). The T test was performed to verify a possible significant difference in the levels of cell proliferation in patients with or without Estriol. The normality was verified by the Shapiro-Wilk test, with data following a normal distribution (W = 0.901 and 0.96, p value > 0.3). The equitable variance was verified by the Levine test, which verified the equitable variance in both groups (F (1) = 0.24, p value = 0.62). A mean positive nucleus of 14 (SD = 10.65) was observed in the group of patients with contraceptive plus placebo and a mean of positive nuclei of 20.81 (SD = 9.88) in the group of patients with contraceptive plus Estriol. The ratio of positive nuclei to total was higher in the estriol group than in the control group. However, it was not possible to verify a significant difference in the two groups (T (14.57) = 1.3, p value = 0.215, d Cohen = 0.53). Cohen’s d pointed to a moderate difference effect (table 4). Table 2. Number of positive, negative and total normal epithelial cell nuclei in immunohistochemical investigation of c-myc antigens in Group 1 (COC + Placebo) (N = 10) Table 3. Number of positive, negative and total normal epithelial cell nuclei in immunohistochemical investigation of c-myc antigens in Group 2 (COC + Estriol) (N = 23) Table 4. Inferential results of the ratio of positive normal epithelial cell nuclei as a function of the control and case groups Citation Format: Osmar Pellegrini, Jr., Afonso Nazário, Angela Flavia L. Waitzberg. Assessment of the proliferative activity of breast tissue after the administration of combined oral hormonal contraceptive, associated or not with estriol [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P3-03-27.

Interleukin-34 deficiency aggravates development of colitis and colitis-associated cancer in mice

Although expression of interleukin (IL)-34 is upregulated in active ulcerative colitis (UC), the molecular function and underlying mechanism are largely unclear. To investigate the function of IL-34 in acute colitis, in a wound healing model and in colitis-associated cancer in IL-34-deficient mice. Colitis was induced by administration of dextran sodium sulfate (DSS), and carcinogenesis was induced by azoxymethane (AOM). Whether the impact of IL-34 on colitis was dependent on macrophages was validated by depletion of macrophages in a murine model. The association between IL-34 expression and epithelial proliferation was studied in patients with active UC. IL-34 deficiency aggravated murine colitis in acute colitis and in wound healing phase. The effect of IL-34 on experimental colitis was not dependent on macrophage differentiation and polarization. IL-34-deficient mice developed more tumors than wild-type mice following administration of AOM and DSS. No significant difference was shown in degree of cellular differentiation in tumors between wild-type and IL-34-deficient mice. IL-34 was dramatically increased in the active UC patients as previously reported. More importantly, expression of IL-34 was positively correlated with epithelial cell proliferation in patients with UC. IL-34 deficiency exacerbates colonic inflammation and accelerates colitis-associated carcinogenesis in mice. It might be served as a potential therapeutic target in UC.

Abstract CT140: ANV419, an IL-2R-beta-gamma targeted antibody-IL-2 fusion protein, induces selective effector cell proliferation in patients with progressed cancer

Abstract Developing a safe and effective non-alpha IL-2R agonist to enhance the numbers and activation status of immune effector cells for the treatment of cancer has remained an elusive goal since the initial approval of Interleukin-2 (IL-2). ANV419 comprises an antibody specific for the IL-2Rα-binding domain of IL-2 fused to native hIL-2 and functions as a potent, highly selective, IL-2Rβγ binding agonist. ANV419 has shown high effector selectivity and a favorable safety profile in preclinical testing, including in non-human primates, and is currently being investigated in an ongoing phase I/II dose finding study in multiple tumor types (ANV419-001). Here we present data from the single patient cohort and the initial 3+3 cohort dose escalation. Data cut-off was December 22nd, 2021. Ten patients in five dose cohorts. Patients had progressed melanoma (cutaneous (n=3), uveal (n=2), mucosal (n=1), choroidal (n=1)), renal cell carcinoma(n=1), hepatocellular carcinoma (n=1) and colorectal cancer(n=1) received Q2W doses of ANV419 of 3mcg/kg (n=1), 6mcg/kg (n=1), 12mcg/kg (n=1), 24mcg/kg (n=4) and 48mcg/kg (n=3) body weight without immunosuppressive premedication as a 15-minute intravenous infusion. Patients received a mean of 4 cycles. Minimal follow up of patients completing the dose limiting toxicity (DLT) period is 2 cycles. All patients experienced mild chills (Grade 1) with or without low-grade fever (Grade 1) hours after drug administration. These resolved with antipyretic treatment within hours. There were two grade 2 AEs in two patients related to ANV419. One patient developed Grade 2 CRS (hypotension grade 2 with fever (Grade 1) and chills (Grade 1) that resolved with 500ml intravenous fluid. One patient experienced transient, self-limiting Grade 2 CRS. No drug related Adverse Events >Grade 2 and no DLTs were observed. One patient in the 24 mcg/kg cohort was non evaluable and was subsequently replaced. Pharmacodynamic evaluation on day 4 post-dosing, showed a dose dependent increase of Ki-67 positivity in CD8 T cells (2%, 14%, 37%, 62%, 62% vs. baseline mean (BLM) 2%)) and NK cells (30%, 62%, 75%, 85%, 80% vs BLM 6%) but not CD4 regulatory T cells (4%, 13%, 25%, 16%, 19% vs. BLM 5%) at 3, 6, 12, 24 and 48 mcg/kg doses respectively. Pharmacokinetic data show a dose proportional increase of serum concentration of ANV419. Both PK and PD were conserved over multiple cycles. Four patients continue to receive ANV419. Four patients underwent more than one follow-up imaging, all of which showed stable disease. Updated patient data will be presented during the meeting. ANV419 is very well tolerated and induces a high degree of proliferation in CD8 T cells and NK cells but not Tregs. ANV419-001 will further evaluate the safety and efficacy of ANV419 alone and in combination with other immunotherapy drugs. Citation Format: Elena Garralda, Alonso Guzman, Juanita Lopez, Vicky Sanchez-Perez, Heinz Läubli, Emiliano Calvo, Christoph Huber, Nicole Egli, Aswathy Nair, Julie Mouton, Kirsten Richter, Carlo Lanza, Sangeeta Jethwa, Silvio Costanzo, Christoph Bucher. ANV419, an IL-2R-beta-gamma targeted antibody-IL-2 fusion protein, induces selective effector cell proliferation in patients with progressed cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr CT140.

Synthetic antigenic determinants of clavulanic acid induce dendritic cell maturation and specific T cell proliferation in patients with immediate hypersensitivity reactions.

Immediate drug hypersensitivity reactions (IDHRs) to clavulanic acid (CLV) have increased in the last decades due to a higher consumption alongside amoxicillin (AX). Due to its chemical instability, diagnostic procedures to evaluate IDHRs to CLV are difficult, and current in vitro assays do not have an optimal sensitivity. The inclusion of the specific metabolites after CLV degradation, which are efficiently recognised by the immune system, could help to improve sensitivity of in vitro tests. Recognition by dendritic cells (DCs) of CLV and the synthetic analogues of two of its hypothesised antigenic determinants (ADs) was evaluated by flow cytometry in 27 allergic patients (AP) and healthy controls (HC). Their ability to trigger the proliferation of T cells was also analysed by flow cytometry. The inclusion of synthetic analogues of CLV ADs, significantly increased the expression of maturation markers on DCs from AP compared to HC. A different recognition pattern could be observed with each AD, and, therefore, the inclusion of both ADs achieves an improved sensitivity. The addition of synthetic ADs analogues increased the proliferative response of CD4+ Th2 compared to the addition of native CLV. The combination of results from both ADs increased the sensitivity of proliferative assays from 19% to 65% with a specificity higher than 90%. Synthetic ADs from CLV are efficiently recognised by DCs with ability to activate CD4+ Th2 cells from AP. The combination of analogues from both ADs, significantly increased the sensitivity of DC maturation and T-cell proliferation compared to native CLV.

Abstract OT2-11-07: Solti-1905. Elacestrant in preoperative setting, a window of opportunity study (ELIPSE trial)

Abstract Background.Patients with hormone receptor-positive/HER2-negative (HR+/HER2-) breast cancer (BC) are mainly treated with therapies that target estrogen receptor (ER) signaling to impair tumor cell proliferation. Several drugs targeting the ER pathway are currently used in the clinical practice including aromatase inhibitors (AI) that limit the production of estradiol, selective ER modulators (SERMs) that compete with estradiol for binding ER, and selective ER degraders (SERDs) that induce ER degradation. Despite the availability of these drugs, finding new ET remains a clinical unmet need since some patients do not initially respond to these treatments or become resistant to available endocrine agents. Elacestrant is an orally bioavailable drug that acts as SERD in breast tissue promoting degradation of ER and inhibiting breast cancer cell proliferation. Several clinical studies have confirmed the tolerable safety profile of elacestrant in BC (Bardia A et al. ASCO, 2021) and a phase III trial investigating elacestrant versus standard ET in metastatic BC patients is ongoing for patients treated with at least 1 prior ET and a CDK4/6 inhibitor for advanced BC (EMERALD; NCT03778931). In this study, we aim to investigate the biological effect of elacestrant on cell proliferation in patients with ER+/HER2- resectable BC. Study design. ELIPSE is prospective, multicenter, single-arm, window-of-opportunity study designed to evaluate the biological effect of elacestrant in treatment naïve patients with ER+/HER2- resectable BC. The study population consists of postmenopausal women with clinically negative axillary lymph node (cN0) and whose primary tumors are≥ 1.5 cm by ultrasound with Ki67 ≥10% locally assessed. Patients will receive 400 mg of elacestrant in monotherapy orally, once a day, continuously. After 4 weeks of treatment, surgery will be performed in accordance with local practice. Two biopsies of the same lesion will be obtained as mandatory: a baseline sample and a surgical sample (tumor core-biopsy is permitted if surgery is not performed after 4 weeks of treatment). Plasma samples for biomarkers will be collected at baseline, surgery and end of study visit. The primary objective is to evaluate the Complete Cell Cycle Arrest rate (defined as Ki67 ≤ 2.7%) by central assessment after 4 weeks of elacestrant therapy. Secondary endpoints include:. 1) safety,. 2) correlation of biological activity determined by changes in: Ki67 as a continuous variable, intrinsic PAM50 subtypes, proliferative signatures, tumor cellularity and tumor-infiltrating lymphocytes (CelTIL, Nuciforo P et al., Ann Oncol, 2018) in tumor samples taken before and at the end of treatment, and. 3) analysis of ctDNA dynamics after treatment. As of July 2021, 7 patients have been enrolled in 4 sites in Spain. NCT04797728. Acknowledgement. This study is financially supported by Radius Pharmaceuticals, Inc. Citation Format: María Vidal, Montserrat Muñoz, Mireia Margeli, Xavier González, Kepa Amillano, Rodrigo Sánchez-Bayona, Fernando Salvador, Tomás Pascual, Aleix Prat, Meritxell Bellet. Solti-1905. Elacestrant in preoperative setting, a window of opportunity study (ELIPSE trial) [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr OT2-11-07.

Biological activities of a recombinant fortilin from Fenneropenaeus merguiensis.

Human Fortilin, an antiapoptotic protein, has also been implicated in several diseases; however, several potential uses of fortilin have also been proposed. Bearing the implications of fortilin in mind, fortilin analog, which has no complication with diseases, is required. Since a recombinant full-length fortilin from Fenneropenaeus merguiensis (rFm-Fortilin (FL)) reported only 44% (3e-27) homologous to human fortilin, therefore the biological activities of the Fm-Fortilin (FL) and its fragments (F2, F12, and F23) were investigated for potential use against HEMA toxicity from filling cement to pulp cell. The rFm-Fortilin FL, F2, 12, and F23 were expressed and assayed for proliferation activity. The rFm-Fortilin (FL) showed proliferation activity on human dental pulp cells (HDPCs) and protected the cells from 2-hydroxy-ethyl methacrylate (HEMA) at 1–20 ng/ml. In contrast, none of the rFm-Fortilin fragments promoted HDPC growth that may be due to a lack of three conserved amino acid residues together for binding with the surface of Rab GTPase for proliferative activity. In addition, rFm-Fortilin (FL) activated mineralization and trend to suppressed production of proinflammatory cytokines, including histamine (at 10 ng/ml) and TNF-α (at 100 ng/ml). Besides, the rFm-Fortilin (FL) did not mutate the Chinese hamster ovary (CHO) cell. Therefore, the rFm-Fortilin (FL) has the potential use as a supplementary medical material to promote cell proliferation in patients suffering severe tooth decay and other conditions.

The prognostic role of circulating CD8+ T cell proliferation in patients with untreated extensive stage small cell lung cancer

BackgroundImmunosuppression caused by tumorigenesis may promote tumor progress and invasion. Here, we investigated whether the characteristics of circulating T lymphocyte subtypes in patients with extensive small cell lung cancer (ED-SCLC) can be used as an alternative marker of tumor progression.MethodsThis study included 36 newly diagnosed ED-SCLC patients before treatment and the patients were followed up. 22 age and sex-matched healthy volunteers were selected as control. The percentages and proliferation potential of T lymphocyte subpopulations from peripheral blood were measured.ResultsCD4+ CD25+ Foxp3+ regulatory T cells (Tregs) were elevated in ED-SCLC patients compared with healthy controls (p = 0.0083). In contrast, the percentages of CD3+ and CD3+CD4+ T cells were significantly lower in SCLC patients (p < 0.001; p = 0.0014). The proliferation (%divided) of CD8+ T cells of SCLC patients was suppressed compared with healthy controls (p = 0.0058), but not of CD4+ T cells (p = 0.1611). Multivariate analyses showed that the %divided of CD8+ T cells is an independent predictor for PFS (HR: 4.342, 95% CI 1.324–14.245; p = 0.015). The percentages of peripheral Tregs and the degree of chemotherapy or radiotherapy induced lymphopenia negatively correlated with the proliferation of CD8+ T cells (p = 0.0225, r = − 0.379; p = 0.0003, r = − 0.464).ConclusionThe present study indicates that SCLC patients have impaired immunity in peripheral blood, and the proliferation potential of circulating CD8+ T cells is a significant predicator for PFS.

LncRNA PCAT19 negatively regulates p53 in non-small cell lung cancer.

The present study aimed to investigate the influence of long non-coding (lnc)RNA prostate cancer associated transcript (PCAT)19 on the progression of non-small cell lung cancer (NSCLC). It was determined that PCAT19 expression was upregulated in NSCLC tissues and also predicted poor patient survival rate. Additionally, p53 expression was downregulated in NSCLC specimens, which was negatively correlated with PCAT19 expression. Moreover, in H1993 NSCLC cells, silencing of the PCAT19 gene led to an increase in the expression of p53, whilst conversely, its overexpression led to p53 downregulation. PCAT19 silencing was associated with the decreased proliferation rate of NSCLC cells, while PCAT19 overexpression led to increased proliferation. In addition, p53 overexpression, achieved through the transfection of a p53 expression vector, attenuated the effects of PCAT19 overexpression on cell proliferation. In conclusion, the present study demonstrated that PCAT19 negatively regulates the p53 tumor-suppression pathway, promoting cancer cell proliferation in patients with NSCLC.

High expression of the transcriptional coactivator TAZ is associated with a worse prognosis and affects cell proliferation in patients with medulloblastoma.

The transcriptional coactivator tafazzin (TAZ) serves pivotal roles in organ development, tumor initiation and tumor progression. However, to the best of our knowledge, the expression of TAZ and its clinical significance in human medulloblastoma have not been defined. The present study aimed to clarify the clinical and biological significance of TAZ expression in human medulloblastoma. Immunohistochemical staining for TAZ was performed with 72 medulloblastoma and three normal brain tissue samples. A high expression level of TAZ was detected in 65.28% of medulloblastoma tissues, whereas low expression was identified in the normal brain tissues. TAZ expression was significantly associated with medulloblastoma recurrence. However, the expression of TAZ was not associated with sex, age, tumor location, tumor maximal diameter and tumor histology. Furthermore, both the overall survival and tumor-free survival rate of patients with high levels of expression of TAZ were shorter compared with those of patients with tumors expressing low levels of TAZ. In univariate and multivariate Cox regression analyses, TAZ expression was identified as a significant prognostic factor for patients with medulloblastoma. Functionally, downregulation of TAZ inhibited the proliferation and tumor formation of medulloblastoma cells and the expression of cell-cycle associated proteins in Daoy cells. In conclusion, high expression of TAZ may serve as a prognostic marker for patients with medulloblastoma and TAZ may be a potential target for medulloblastoma therapy.

PI3K (Phosphatidylinositol 3-Kinase) Activation and Endothelial Cell Proliferation in Patients with Hemorrhagic Hereditary Telangiectasia Type 1.

Hemorrhagic hereditary telangiectasia (HHT) type 2 patients have increased activation of the phosphatidylinositol 3-kinase (PI3K) signaling pathway in telangiectasia. The main objective is to evaluate the activation of the PI3K pathway in cutaneous telangiectasia of HHT1 patients. A cutaneous biopsy of a digital hand telangiectasia was performed in seven HHT1 and eight HHT2 patients and compared with six controls. The study was approved by the Clinical Research Ethics Committee of our center. A histopathological pattern with more dilated and superficial vessels that pushed up the epidermis was identified in HHT patients regardless of the type of mutation and was associated with older age, as opposed to the common telangiectasia pattern. The mean proliferation index (Ki-67) was statistically higher in endothelial cells (EC) from HHT1 than in controls. The percentage of positive EC for pNDRG1, pAKT, and pS6 in HHT1 patients versus controls resulted in higher values, statistically significant for pNDRG1 and pS6. In conclusion, we detected an increase in EC proliferation linked to overactivation of the PI3K pathway in cutaneous telangiectasia biopsies from HHT1 patients. Our results suggest that PI3K inhibitors could be used as novel therapeutic agents for HHT.

The Correlation Between [68Ga]DOTATATE PET/CT and Cell Proliferation in Patients With GEP-NENs.

Objectives of the study are to analyze the correlation between [68Ga]DOTATATE positron emission tomography (PET)/X-ray computed tomography (CT) measurements and various biological characteristics of gastroenteropancreatic neuroendocrine neoplasms (GEP-NENs), and to determine optimal cutoff value of SUVmax (standard uptake value) to differentiate neuroendocrine tumors (NETs) and neuroendocrine cancers (NECs). Of the GEP-NEN cases (73 males, 53 females; age 18-77years) with pathologically proven primary and/or metastatic lesions, 126 were studied. All of the short axes of lesions were larger than 0.5cm in order to avoid the partial volume effect. Patients fasted for 6h before the PET/CT scans. The dose of [68Ga]DOTATATE was 100-200MBq and the acquisition began at 1h after injection. The lesion with the highest SUVmax in each patient was analyzed. In the total sample, the sensitivity of [68Ga]DOTATATE was 69.05%. The sensitivities were significantly different among G1, G2, and G3 groups (72.22%, 91.53%, and 40.82%, respectively; p < 0.01). The SUVmax of the G3 group was lowest. We also found that the sensitivity and SUVmax were significantly higher (p < 0.05) in patients with pancreatic NENs (Pan-NENs) than in patients with gastrointestinal NENs (Gi-NENs) and unknown primary NENs (Up-NENs). A significant negative correlation between SUVmax and Ki-67 was found (r = - 0.429, p < 0.01). Using SUVmax to differentiate neuroendocrine tumors (NETs) and neuroendocrine cancers (NECs), the area under the ROC curve (AUC) was 0.771 and the cutoff value of SUVmax was 11.25 (sensitivity 79.2%, specificity 65.3%). However, Pan-NENs did not show any statistical significance results in correlation and ROC analysis. [68Ga]DOTATATE PET/CT results showed a negative correlation with GEP-NEN cell proliferation and were complementary to Ki-67. Pan-NENs were different from Gi-NENs and Up-NENs when compared to somatostatin receptor expression.

Abstract P1-15-25: Prognostic factors associated with pathological complete response in early breast cancer patients undergoing neoadjuvant chemotherapy. The importance of Ki-67 and molecular subtype

Abstract Background: Ki-67 immunohistochemical determination is a widely used biomarker of cell proliferation in patients (pts) undergoing endocrine treatment for breast BC. The role of Ki-67 in pts undergoing neoadjuvant chemotherapy (NAC) for early BC remains controversial. Methods: We analyzed retrospectively data on 137 patients undergoing taxane and/or anthracycline, transtuzumab based NAC. Luminal A was documented in 6 pts, Luminal B in 29 pts, Her-2 positive in 30 pts and triple negative breast cancers (TNBC) in 72 pts. Pathological complete response (pCR) was defined as the complete disappearance of the invasive cancer in the breast and absence of tumor in the axillary lymph nodes examined by axillary clearance. Results: The pCR rate of the entire cohort was 41.6%. At 2 years 92% of pts who attained a pCR were disease free compared to 80% of pts who did not attain a pCR (log rank test p &amp;lt; 0.0147). On univariate analysis factors associated with higher pCR included primary tumor size (T1 68% vs. T2 41% vs. T3 or T4 0%, Chi2=20.05, p&amp;lt;0.00017), nodal disease (N0 49% vs. N1 39% vs. N2 8%, p&amp;lt;0.02948), ER receptor status (negative 59% vs. positive 14%, p&amp;lt;0.00000), PR receptor status (negative 53% vs. positive 17%, p&amp;lt;0.00002), molecular subtype (TNBC 53.4%, Her2=50% and Luminal A + B was 8.5%, p&amp;lt;0.00002), Ki67 (&amp;gt;40=55% vs. 15-39=34% vs. &amp;lt;15=0%, p&amp;lt;0.00060) and Stage (I= 85% vs. IIA=49% vs. IIB=36% vs. III=5%, p&amp;lt;0.00006). Factors not associated with a higher pCR included age, menopausal status, extranodal spread and lympho-vascular invasion. In a logistic regression model Ki-67 as a continuous variable (p&amp;lt;0.01203) and molecular subtype (p&amp;lt;0.02228) retained its significance; while tumor size, stage of disease, nodal status, ER and PR loss significance. Conclusion: Ki67 and molecular subtype (Her-2 positive disease and TNBC) are independent prognostic factors of pCR in pts with early BC undergoing NAC. Citation Format: Rapoport BL, Barnard-Tide J, Smit T, Nayler S, Benn C-A. Prognostic factors associated with pathological complete response in early breast cancer patients undergoing neoadjuvant chemotherapy. The importance of Ki-67 and molecular subtype [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P1-15-25.

Expression of T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain on CD4+ T cells in patients with atopic dermatitis.

The T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain (TIGIT) is a co-inhibitory receptor mainly expressed on T cells. Although TIGIT plays an important role in various autoimmune diseases, its role in atopic dermatitis (AD) remains unclear. In this study, we examined the expression levels of TIGIT and their association with clinical features in patients with AD. TIGIT expression on CD4+ T cells, central memory T cells, effector memory T cells and regulatory T cells was determined by flow cytometry. CD4+ T cells exhibited enhanced TIGIT expression in patients with AD compared with healthy individuals. In particular, effector memory T cells and regulatory T cells, but not central memory T cells, exhibited higher TIGIT expression in patients with AD than in healthy individuals. The frequency of TIGIT+ cells among CD4+ T cells was significantly increased in patients with mild AD compared with healthy individuals, while decreased in patients with severe AD. Consistently, the frequency of TIGIT+ cells among CD4+ T cells was negatively correlated with both serum thymus and activation-regulated chemokine levels and immunoglobulin E levels in patients with AD. Furthermore, TIGIT expression on CD4+ T cells inhibited cell proliferation in patients with AD. These results suggest that TIGIT expression on CD4+ T cells in patients with AD may be increased to suppress chronic cutaneous inflammation. Moreover, TIGIT expression may be impaired in a subset of patients with AD, leading to a deterioration of skin inflammation. Our study may provide new insight into a TIGIT pathway-based therapeutic approach for AD.

Erratum: Mismatched intratumoral distribution of [18F] fluorodeoxyglucose and 3'-deoxy-3'-[18F] fluorothymidine in patients with lung cancer.

[This corrects the article DOI: 10.3892/ol.2017.6840.].

Overactivation of Ca2+/Calmodulin‐Dependent Protein Kinase IV and IIδ Contributes to Enhancing Pulmonary Arterial Smooth Muscle Cell Proliferation in Patients with Idiopathic Pulmonary Arterial Hypertension

IntroductionA rise in cytosolic free Ca2+ concentration ([Ca2+]cyt) in pulmonary arterial smooth muscle cells (PASMCs) causes pulmonary vasoconstriction and increases PASMC proliferation, leading to concentric arterial wall thickening. Ca2+/Calmodulin‐Dependent Protein Kinases (CaMK) are a family of serine/threonine kinases that are activated by an increase in [Ca2+]cyt. Akt is another serine/threonine protein kinase that has been observed to have increased activity due to a rise in [Ca2+]cyt, and its overexpression has been implicated in excessive cell proliferation. CaMKIV has been shown to cause phosphorylation of Akt while CaMKIIδ has been shown to cause increases in vascular remodeling and vascular smooth muscle cell proliferation. STIM2, a Ca2+ sensor in sarcoplasmic reticulum (SR), has been suggested to mediate increases in the resting [Ca2+]cyt in PASMCs from patients with idiopathic pulmonary arterial hypertension (IPAH). PDGF‐R is a tyrosine kinase receptor that, when activated by PDGF, can cause cell proliferation. Our hypothesis is that overactivation of CaMKIV and CaMKIIδ increases activity of Akt, causing upregulation of STIM2 and PDGF‐R, which stimulate PASMC proliferation in patients with IPAH.MethodsPASMCs from both healthy subjects and patients with IPAH were used for this study. Measurement of protein levels was done using conventional western blot protocol. Knockdown of specific proteins was done using siRNA. Measurement of cell proliferation was done by measuring EdU incorporation rate, an indicator of DNA synthesis, and total cell number after 48 hours.ResultsIn this study, we found that the protein level of CaMKIV is significantly increased in PASMCs from patients with IPAH (1.00±0.588 vs 11.4±1.29, n=3 normal subjects and 5 patients, p=0.001). Knockdown of CaMKIV in IPAH PASMCs with CaMK IV specific siRNA significantly decreased the protein level of STIM2, PDGF‐R‐α and PDGF‐R‐β, and also significantly decreased cell proliferation, which is indicated by the decreased EdU incorporation rate and total cell number in IPAH PASMCs. Knockdown of CaMKIIδ significantly decreased phosphorylation of Akt, EdU incorporation rate and total cell number. In addition, knockdown of Akt resulted in a decrease in STIM2 and PDGF‐R‐β.ConclusionTogether, these data indicate that overactivation of CaMKIV and CaMKIIδ in IPAH PASMCs, due to upregulation of CaMKIV, CaMKIIδ and/or increased resting [Ca2+]cyt, contributes to enhancing PASMCs proliferation by increasing activation of Akt, causing upregulation of STIM2 and PDGF‐R. CaMKIV and CaMKIIδ activation‐induced upregulation of STIM2 will further raise the resting [Ca2+]cyt and subsequently activate CaMKIV and CaMKIIδ, forming a positive feedback loop. The overactivation of CaMKIV and CaMKIIδ may be an important mechanism involved in sustained pulmonary vasoconstriction and excessive pulmonary vascular remodeling in patients with IPAH.Support or Funding InformationThis work was supported in part by the grants from the National Heart, Lung and Blood Institute of the National Institutes of Health (HL135807 and HL125208).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Autophagy, Apoptosis, and Cell Proliferation in Exstrophy-Epispadias Complex

To investigate the state of autophagy and its interactions with apoptosis and cell proliferation in patients who underwent successful early closure or delayed closure of exstrophy. They compared those outcomes with cell culture samples from patients with vesicoureteral reflux as control. Primary cultures of bladder smooth muscle cells (SMCs) were established from patients with successful neonatal bladder closure (group 1, N = 5), delayed closure because of small bladder template (group 2, N = 5), and vesicoureteral reflux as control (group 3, N = 5). The myogenicity of the cultures was determined using anti-Desmin antibody. Immunostainings for LC3 to assess autophagy and Ki67 to assess cell proliferation were applied. Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated dUTP digoxigenin nick-end labeling assay. Autophagy marker (LC3) expression was significantly higher in the delayed closure group than in the other groups, whereas no significant difference was noted between the neonatal closure and the control groups. Apoptotic indices of the SMCs were remarkably higher in SMC cultures from the delayed closures than in the neonatal closure and the control groups. A significantly lower expression of proliferation marker (Ki67) in the delayed closure group compared with the control and the neonatal closure group was also of note. Patients with small bladder template and delayed closure showed upregulated autophagic process and increased apoptotic indices while experiencing a dramatic decrease in the proliferation of their bladder SMCs. Finally, the concept of manipulating autophagy may lead to promising outcomes for patients with bladder exstrophy in the future.

Mismatched intratumoral distribution of [18F] fluorodeoxyglucose and 3'-deoxy-3'-[18F] fluorothymidine in patients with lung cancer.

In a mouse model of human lung cancer, intratumoral distribution between 3′-deoxy-3′-[18F] fluorothymidine (18F-FLT) and [18F] fluorodeoxyglucose (18F-FDG) was mutually exclusive. 18F-FLT primarily accumulated in proliferating cancer cells, whereas 18F-FDG accumulated in hypoxic cancer cells. The aim of the present study was to evaluate these preclinical findings in patients with lung cancer. A total of 55 patients with solitary pulmonary lesion were included in the present study. Patients underwent 18F-FLT positron emission tomography-computed tomography (PET/CT) and 18F-FDG PET/CT scan with a 3-day interval. The final diagnosis was based on histological examination. Among the 55 cases, a total of 24 cases were confirmed as malignant lesions. Mismatched 18F-FLT- and 18F-FDG-accumulated regions were observed in 19 cases (79%) and matched in 5 (21%). Among the 31 benign lesions, 18F-FLT and 18F-FDG were mismatched in 12 cases (39%) and matched in 19 (61%). The difference in intratumoral distribution of 18F-FLT and 18F-FDG between malignant and benign lesions was statistically significant (P<0.05). The results of the present study indicate that a mismatch in intratumoral distribution of 18F-FLT and 18F-FDG may be a feature of patients with lung cancer. Increased 18F-FDG accumulation may serve as an indicator of tumor hypoxia, whereas regions with increased 18F-FLT uptake may be associated with an increased rate of cancer cell proliferation in patients with lung cancer.

Abstract P5-04-02: Serum thymidine kinase 1 activity as a pharmacodynamics marker of cyclin-dependent kinase 4/6 inhibition in patients with early stage breast cancer receiving neoadjuvant palbociclib

Abstract Background: Thymidine kinase 1 (TK1) is a fundamental enzyme in DNA synthesis. TK1 expression is E2F-dependent and peaks in the S-phase of the cell cycle. In preclinical studies, inhibition of cyclin-dependent kinase (CDK) 4/6 led to dose dependent reduction of TK1 activity in cultured media. We hypothesized that serum TK1 could serve as a non-invasive surrogate marker of cell proliferation in patients (pts) receiving CDK4/6 inhibitors. In this study, we examined serum TK1 activity from breast cancer (BC) pts enrolled on a neoadjuvant study of palbociclib (Palbo) plus anastrozole (A), for changes induced by Palbo, and correlated with changes in tumor Ki67. Methods: In this phase II neoadjuvant study, 50 pts with clinical stage II or III estrogen receptor positive (ER+) HER2- BC, received A (in combination with goserelin if premenopausal) alone for 28 days in cycle 0 (C0), followed by the addition of Palbo (125 mg daily on days 1-21) on cycle 1 day 1 (C1D1) for 4 28-day cycles (C1 to C4) unless C1D15 tumor Ki67&amp;gt;10%, in which case pts went off study. Following completion of cycle 4, A was continued for another 3-5 weeks to allow Palbo washout prior to surgery, except in 8 pts who received an additional 10-12 days of Palbo immediately prior. Blood and tumor biopsies were collected at 4 time points: baseline, C1D1, C1D15, and surgery. Serum TK1 activity was measured using the highly sensitive Divitum™ assay according to the Divitum™ Instructions for use (Biovica, Sweden). Tumor Ki67 IHC was performed at the Washington University AMP laboratory using the CONFIRM anti-Ki67 rabbit monoclonal antibody (clone 30-9), and pathologist guided image analysis. Results: There was no statistically significant difference in TK activity between baseline and C1D1 serum samples (Table 1). However, serum TK activity decreased significantly from C1D1 to C1D15 following the addition of Palbo and increased significantly from C1D15 to surgery following Palbo washout (Table 1), indicating a significant effect of Palbo on TK activity. At C1D15, TK activity was below the detection limit of 20 Du/L in 44 of 48 pts, and was at low levels (24, 26, 26, and 58 Du/L) in the remaining 4 pts, indicating a profound effect by Palbo. Interestingly, the TK activities of the 4 pts with tumor Ki67 &amp;gt;10% at C1D15 were all below 20 Du/L, suggesting the possibility of tumor cell proliferation independent of CDK4/6 inhibition. The sensitivity and specificity of change (increase/decrease) in serum TK activity to predict tumor Ki67 (increase/decrease) induced by Palbo were 83% (19/23, 95%CI: 66-99%) and 93% (26/28, 95%CI: 83%-100%), respectively. The Kappa statistic was 0.761 (P&amp;lt;0.001), indicating substantial agreement between the two tests. Conclusions: Serum TK1 activity may serve as a pharmacodynamics marker of CDK4/6 inhibition and further investigation is warranted. Table 1. Serum TK1 and tumor Ki67 Serum TKKi67 Median (IQR) (Du/L)NMedian (IQR) (%)NBaseline46 (25-73)4824.34% (11.92%-35.43%)45Cycle 1 day 143 (27.5-98)495.37% (2.49%-13.59%)*45Cycle 1 day 1520 (20-20)*480.78% (0.23%-1.05%)*45Day of surgery136.0 (37.5-259)*378.33% (2.25%-23.03%)*34*P&amp;lt;0.001 compared to the preceding time point. Citation Format: Liu N, Thomas S, Luo R, Hoog J, Suh EM, Bergqvist M, Neumüller M, Guo Z, Vij K, Sanati S, Ellis M, Ma C. Serum thymidine kinase 1 activity as a pharmacodynamics marker of cyclin-dependent kinase 4/6 inhibition in patients with early stage breast cancer receiving neoadjuvant palbociclib [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P5-04-02.