bluuebunny/crossref_metadata_2025 huggingface dataset:
Testing an infection model to explain excess risk of preterm birth with long-term iron supplementation in a malaria endemic area
Authors: Brabin Bernard , Tinto Halidou , Roberts Stephen A.
Published: November 2019

Background

In view of recent evidence from a randomized trial in Burkina Faso that periconceptional iron supplementation substantially increases risk of spontaneous preterm birth (< 37 weeks) in first pregnancies (adjusted relative risk = 2.22; 95% CI 1.39–3.61), explanation is required to understand potential mechanisms, including progesterone mediated responses, linking long-term iron supplementation, malaria and gestational age.

Methods

The analysis developed a model based on a dual hit inflammatory mechanism arising from simultaneous malaria and gut infections, supported in part by published trial results. This model is developed to understand mechanisms linking iron supplementation, malaria and gestational age. Background literature substantiates synergistic inflammatory effects of these infections where trial data is unavailable. A path modelling exercise assessed direct and indirect paths influencing preterm birth and gestation length.

Results

A dual hit hypothesis incorporates two main pathways for pro-inflammatory mechanisms, which in this model, interact to increase hepcidin expression. Trial data showed preterm birth was positively associated with C-reactive protein (P = 0.0038) an inflammatory biomarker. The malaria pathway upregulates C-reactive protein and serum hepcidin, thereby reducing iron absorption. The enteric pathway results from unabsorbed gut iron, which induces microbiome changes and pathogenic gut infections, initiating pro-inflammatory events with lipopolysaccharide expression. Data from the trial suggest that raised hepcidin concentration is a mediating catalyst, being inversely associated with shorter gestational age at delivery (P = 0.002) and positively with preterm incidence (P = 0.007). A segmented regression model identified a change-point consisting of two segments before and after a sharp rise in hepcidin concentration. This showed a post change hepcidin elevation in women with increasing C-reactive protein values in late gestation (post-change slope 0.55. 95% CI 0.39–0.92, P < 0.001). Path modelling confirmed seasonal malaria effects on preterm birth, with mediation through C-reactive protein and (non-linear) hepcidin induction.

Conclusions

Following long-term iron supplementation, dual inflammatory pathways that mediate hepcidin expression and culminate in progesterone withdrawal may account for the reduction in gestational age observed in first pregnancies in this area of high malaria exposure. If correct, this model strongly suggests that in such areas, effective infection control is required prior to iron supplementation to avoid increasing preterm births.

Trial registration NCT01210040. Registered with Clinicaltrials.gov on 27th September 2010
Microglial VPS35 deficiency regulates microglial polarization and decreases ischemic stroke-induced damage in the cortex
Authors: Ye Shi-Yang , Apple Joanna E. , Ren Xiao , Tang Fu-Lei , Yao Ling-Ling , Wang Yong-Gang , Mei Lin , Zhou Yuan-Guo , Xiong Wen-Cheng
Published: November 2019

Background

Vacuolar sorting protein 35 (VPS35), a critical component of retromer, is essential for selective endosome-to-Golgi retrieval of membrane proteins. It is highly expressed in microglial cells, in addition to neurons. We have previously demonstrated microglial VPS35’s functions in preventing hippocampal, but not cortical, microglial activation, and in promoting adult hippocampal neurogenesis. However, microglial VPS35’s role in the cortex in response to ischemic stroke remains largely unclear.

Methods

We used mice with VPS35 cKO (conditional knockout) in microglial cells and examined and compared their responses to ischemic stroke with control mice. The brain damage, cell death, changes in glial cells and gene expression, and sensorimotor deficits were assessed by a combination of immunohistochemical and immunofluorescence staining, RT-PCR, Western blot, and neurological functional behavior tests.

Results

We found that microglial VPS35 loss results in an increase of anti-inflammatory microglia in mouse cortex after ischemic stroke. The ischemic stroke-induced brain injury phenotypes, including brain damage, neuronal death, and sensorimotor deficits, were all attenuated by microglial VPS35-deficiency. Further analysis of protein expression changes revealed a reduction in CX3CR1 (CX3C chemokine receptor 1) in microglial VPS35-deficient cortex after ischemic stroke, implicating CX3CR1 as a potential cargo of VPS35 in this event.

Conclusion

Together, these results reveal an unrecognized function of microglial VPS35 in enhancing ischemic brain injury-induced inflammatory microglia, but suppressing the injury-induced anti-inflammatory microglia. Consequently, microglial VPS35 cKO mice exhibit attenuation of ischemic brain injury response.
BRCA1 mutation influences progesterone response in human benign mammary organoids
Authors: Davaadelger Batzaya , Choi Mi-Ran , Singhal Hari , Clare Susan E. , Khan Seema A. , Kim J. Julie
Published: November 2019

Background

Women, who carry a germline BRCA1 gene mutation, have a markedly increased risk of developing breast cancer during their lifetime. While BRCA1 carriers frequently develop triple-negative, basal-like, aggressive breast tumors, hormone signaling is important in the genesis of BRCA1 mutant breast cancers. We investigated the hormone response in BRCA1-mutated benign breast tissue using an in vitro organoid system.

Methods

Scaffold-free, multicellular human breast organoids generated from benign breast tissues from non-carrier or BRCA1 mutation carriers were treated in vitro with a stepwise menstrual cycle hormone regimen of estradiol (E2) and progesterone (P4) over the course of 28 days.

Results

Breast organoids exhibited characteristics of the native breast tissue, including expression of hormone receptors, collagen production, and markers of luminal and basal epithelium, and stromal fibroblasts. RNA sequencing analysis revealed distinct gene expression in response to hormone treatment in the non-carrier and BRCA1-mutated organoids. The selective progesterone receptor modulator, telapristone acetate (TPA), was used to identify specifically PR regulated genes. Specifically, extracellular matrix organization genes were regulated by E2+P4+TPA in the BRCA1-mutated organoids but not in the non-carrier organoids. In contrast, in the non-carrier organoids, known PR target genes such as the cell cycle genes were inhibited by TPA.

Conclusions

These data show that BRCA1 mutation influences hormone response and in particular PR activity which differs from that of non-carrier organoids. Our organoid model system revealed important insights into the role of PR in BRCA1-mutated benign breast cells and the critical paracrine actions that modify hormone receptor (HR)-negative cells. Further analysis of the molecular mechanism of BRCA1 and PR crosstalk is warranted using this model system.
Low coverage whole genome sequencing enables accurate assessment of common variants and calculation of genome-wide polygenic scores
Authors: Homburger Julian R. , Neben Cynthia L. , Mishne Gilad , Zhou Alicia Y. , Kathiresan Sekar , Khera Amit V.
Published: November 2019

Background

Inherited susceptibility to common, complex diseases may be caused by rare, pathogenic variants (“monogenic”) or by the cumulative effect of numerous common variants (“polygenic”). Comprehensive genome interpretation should enable assessment for both monogenic and polygenic components of inherited risk. The traditional approach requires two distinct genetic testing technologies—high coverage sequencing of known genes to detect monogenic variants and a genome-wide genotyping array followed by imputation to calculate genome-wide polygenic scores (GPSs). We assessed the feasibility and accuracy of using low coverage whole genome sequencing (lcWGS) as an alternative to genotyping arrays to calculate GPSs.

Methods

First, we performed downsampling and imputation of WGS data from ten individuals to assess concordance with known genotypes. Second, we assessed the correlation between GPSs for 3 common diseases—coronary artery disease (CAD), breast cancer (BC), and atrial fibrillation (AF)—calculated using lcWGS and genotyping array in 184 samples. Third, we assessed concordance of lcWGS-based genotype calls and GPS calculation in 120 individuals with known genotypes, selected to reflect diverse ancestral backgrounds. Fourth, we assessed the relationship between GPSs calculated using lcWGS and disease phenotypes in a cohort of 11,502 individuals of European ancestry.

Results

We found imputation accuracy r2 values of greater than 0.90 for all ten samples—including those of African and Ashkenazi Jewish ancestry—with lcWGS data at 0.5×. GPSs calculated using lcWGS and genotyping array followed by imputation in 184 individuals were highly correlated for each of the 3 common diseases (r2 = 0.93–0.97) with similar score distributions. Using lcWGS data from 120 individuals of diverse ancestral backgrounds, we found similar results with respect to imputation accuracy and GPS correlations. Finally, we calculated GPSs for CAD, BC, and AF using lcWGS in 11,502 individuals of European ancestry, confirming odds ratios per standard deviation increment ranging 1.28 to 1.59, consistent with previous studies.

Conclusions

lcWGS is an alternative technology to genotyping arrays for common genetic variant assessment and GPS calculation. lcWGS provides comparable imputation accuracy while also overcoming the ascertainment bias inherent to variant selection in genotyping array design.
The intestinal microbial community dissimilarity in hepatitis B virus-related liver cirrhosis patients with and without at alcohol consumption
Authors: Deng Yong-Dong , Peng Xue-Bin , Zhao Rong-Rong , Ma Chao-Qun , Li Jian-ning , Yao Li-Qiong
Published: November 2019

Background

Chronic hepatitis B virus (HBV) infection-reduced liver functions are associated with intestinal microbial community dissimilarity. This study aimed to investigate the microbial community dissimilarity in patients with different grades of HBV-related liver cirrhosis.

Results

Serum endotoxin was increased with Child–Pugh (CP) class (A, B, and C). Veillonellaceae and Lachnospiraceae families were reduced in patients compared with controls. Megamonas and Veillonella genus was reduced and increased in patients compared with controls, respectively, especially in CPB and CPC groups. Correlation analysis showed that endotoxin content was significantly correlated with alcohol consumption (95% CI 0.100, 0.493), CP class (95% CI 0.289, 0.687) and Lachnospiraceae family level (95% CI − 0.539, − 0.122). Firmicutes/Bacteroidetes ratio was correlated with the level of Lachnospiraceae family (95% CI 0.013, 0.481), Veillonellaceae family (95% CI 0.284, 0.696), Megamonas genus (95% CI 0.101, 0.518) and Veillonella genus (95% CI 0.134, 0.545). All aforementioned bacteria were independent risk or protective factors for hepatitis. Alcohol consumption changed microbial community.

Conclusions

Our study demonstrated that elevated Firmicutes/Bacteroidetes ratio, reduced Megamonas genus level and increased Veillonella genus level were indicators for HBV-related liver cirrhosis. Alcohol-related pathogenesis was associated with the changed microbial community.
Short-term skin problems in infants aged 0–3 months affect food allergies or atopic dermatitis until 2 years of age, among infants of the general population
Authors: Yonezawa Kaori , Haruna Megumi
Published: November 2019

Background

This study examined whether infants aged 0–3 months exhibited long-term effects of using a moisturizer skincare intervention and whether a short-term skin problem resulted in the subsequent development of food allergies or atopic dermatitis (AD) until the age of 2 years.

Methods

This study was a follow-up of a completed randomized control trial (RCT) of moisturizer skincare for infants aged 0–3 months. A self-reported questionnaire was mailed to the parents of children aged 1–2 years who had participated in the RCT. Data were analyzed using a Chi square test, by intention to treat analysis, and by multiple logistic regression.

Results

Of 155 infants, 22 (14.2%) and 28 (18.1%) had food allergies and AD/eczema until 2 years of age, respectively. No significant difference was seen in food allergies or AD between the group that received moisturizer skincare intervention and the control group. On the contrary, food allergies until 2 years of age were significantly associated with short-term (4–7 days) and long-term (more than 7 days) body skin problems occurring in the first 3 months of life, a family history of AD, and the time of starting complementary food. High value of face transepidermal water loss at 3 months of age was also associated with food allergies. Moreover, a short duration of severe diaper dermatitis during the first 3 months, a family history of AD, and being male were significantly associated with AD/eczema until the age of 2 years.

Conclusions

After adjusting for family history of AD, a short-term skin problem in the first 3 months of life was significantly associated with the development of food allergies or AD/eczema until the age of 2 years. Prevention or prompt treatment of skin problems in newborns is essential for preventing future allergic diseases.

Trial registration This was a follow-up study conducted 2 years after the completed RCT of a moisturizer skincare intervention for early infants, which was registered in the University Hospital Medical Information Network Clinical Trials Registry (UMIN000013260)
Use of a vibrating mesh nebulizer for allergen challenge
Authors: Cockcroft Donald W. , Davis Beth E. , Blais Christianne M. , Boulet Louis-Philippe , Boulay Marie-Éve , Villeneuve Hélène , Gauvreau Gail M. , O’Byrne Paul M. , Howie Karen J. , Obminski Caitlin D.
Published: November 2019

Background

Allergen inhalation tests are a valuable research tool. The allergen dose producing an early asthmatic response (EAR) can be predicted from methacholine responsiveness and allergen skin test endpoint (STE). The Wright® jet nebulizer, which is both inefficient and increasingly difficult to obtain, has been used historically. We assessed the Solo® vibrating mesh nebulizer as an alternative for allergen and methacholine challenges.

Methods

Eighteen mild atopic asthmatics completed the study. Doubling concentration allergen prick skin tests were performed to determine the STE in allergen units/mL. The Wright® protocol was used to measure the methacholine provocation dose causing a 20% forced expired volume in one second (FEV1) fall (PD20) (μg) and the allergen PD20 (units). The Solo® protocol (0.5 mL nebulized to completion, tidal breathing inhalation) was used to determine both methacholine PD20 and allergen PD20. The nebulizer order was randomized and separated by ≥ 2 weeks.

Results

All data were log transformed. The allergen PD20, predicted from the methacholine PD20 and the STE, was within 2 doubling doses of the PD20 measured with the Wright® and 2.64 doubling doses of that measured with Solo®. The Wright® allergen PD20 correlated with the Wright® methacholine PD20 (r = 0.74) and the STE (r = 0.78) and more strongly with the product of the two (Wright® methacholine PD20 × STE, r = 0.91, p < 0.00001). The Solo® allergen PD20 showed similar relationships with the Solo® methacholine PD20 (r = 0.61), the STE (r = 0.75) and the product of the two (Solo® methacholine PD20 × STE, r = 0.83, p < 0.00002). The Wright® and the Solo® methacholine geometric mean PD20s were not significantly different (49.3 and 54.5 μg respectively, p = 0.62). The Wright® allergen PD20 was slightly but significantly lower than the Solo® allergen PD20 (geometric means 6.7 and 10.5 units respectively, p = 0.003).

Conclusion

The Solo® allergen PD20 showed the same relationship with methacholine responsiveness and STE as did the Wright®. The Solo® allergen PD20 was slightly but significantly higher than the Wright® allergen PD20. The Solo® vibrating mesh nebulizer was well tolerated and is an acceptable alternative for allergen challenge.

Trial registration clinicaltrials.gov: NCT03491358
Stem cell-derived extracellular vesicles: role in oncogenic processes, bioengineering potential, and technical challenges
Authors: Ullah Mujib , Qiao Yang , Concepcion Waldo , Thakor Avnesh S.
Published: November 2019

Extracellular vesicles (EVs) are cellular-derived versatile transporters with a specialized property for trafficking a variety of cargo, including metabolites, growth factors, cytokines, proteins, lipids, and nucleic acids, throughout the microenvironment. EVs can act in a paracrine manner to facilitate communication between cells as well as modulate immune, inflammatory, regenerative, and remodeling processes. Of particular interest is the emerging association between EVs and stem cells, given their ability to integrate complex inputs for facilitating cellular migration to the sites of tissue injury. Additionally, stem cell-derived EVs can also act in an autocrine manner to influence stem cell proliferation, mobilization, differentiation, and self-renewal. Hence, it has been postulated that stem cells and EVs may work synergistically in the process of tissue repair and that dysregulation of EVs may cause a loss of homeostasis in the microenvironment leading to disease. By harnessing the property of EVs for delivery of small molecules, stem cell-derived EVs possess significant potential as a platform for developing bioengineering approaches for next-generation cancer therapies and targeted drug delivery methods. Although one of the main challenges of clinical cancer treatment remains a lack of specificity for the delivery of effective treatment options, EVs can be modified via genetic, biochemical, or synthetic methods for enhanced targeting ability of chemotherapeutic agents in promoting tumor regression. Here, we summarize recent research on the bioengineering potential of EV-based cancer therapies. A comprehensive understanding of EV modification may provide a novel strategy for cancer therapy and for the utilization of EVs in the targeting of oncogenic processes. Furthermore, innovative and emerging new technologies are shifting the paradigm and playing pivotal roles by continually expanding novel methods and materials for synthetic processes involved in the bioengineering of EVs for enhanced precision therapeutics.
Correction to: Human-as-a-security-sensor for harvesting threat intelligence
Authors: Vielberth Manfred , Menges Florian , Pernul Günther
Published: November 2019

In the original publication of this article (Vielberth et al. 2019), the author list was not completed.
The Parliamentary Scholar Scheme: a way to engage doctors in healthcare policy and politics
Authors: Perry Jen , Lomax Paul , Taylor Fiona , Howson Susan , McCurdy Kathleen
Published: November 2019

The Royal College of Psychiatrists’ Parliamentary Scholar Scheme gives higher trainees in psychiatry the opportunity to spend 1 day a week in the House of Lords working alongside a peer with an interest in health. This article describes the work of the House of Lords and Parliament using examples from the experiences of 2017–2018 scholars and outlines ways doctors can get more involved in policy and politics.
Mutational load (ML) and T-cell-inflamed microenvironment as predictors of response to pembrolizumab.
Authors: Cristescu Razvan , Mogg Robin , Ayers Mark , Albright Andrew , Murphy Erin , Yearley Jennifer , Sher Xinwei , Liu Xiaoqiao , Nebozhyn Michael , Lunceford Jared K. , Joe Andrew K. , Cheng Jonathan , Plimack Elizabeth R. , Ott Patrick Alexander , Seiwert Tanguy Y. , McClanahan Terrill K , Loboda Andrey , Kaufman David Ross
Published: November 2019

1

Background: ML is associated with response to anti CTLA-4 and PD-1/L1 immunotherapies in some cancers, likely due to increased tumor antigenicity via generation of neoepitopes not subject to central immune tolerance. An IFNγ centric gene expression profile (GEP), characteristic of a T cell inflamed microenvironment, is also related to response to PD-1/PD-L1 therapies. The association between ML and outcome, and independent predictive values of ML and GEP in pembrolizumab treated patients across 21 tumor types were assessed. Methods: Whole exome and RNA sequencing were performed on FFPE specimens pooled from 2 multicohort advanced solid tumor trials (KEYNOTE 012 [n = 39] and 028 [n = 80]). The previously defined GEP score is a weighted sum of normalized expression values for 18 genes. ML and neoantigen analysis tools included GATK, MuTect and NetMHC. Statistical testing was adjusted for multiple testing. ML and GEP association was also explored in Moffitt and TCGA datasets. Results: ML and neoantigen load were significantly associated with objective response (OR) (AUROC = 0.76 and 0.78; p = 0.0036 and 0.0083, respectively). Median numbers of mutations were 180 in responders and 61 in nonresponders. The overall response rate (ORR) in all patients was 15%. At a ML cutoff of 102 (based on ROC Curve YoudenIndex), the ORR was 32.3% (prevalence 31.0%; NPV 92.8%). GEP was also significantly associated with OR (AUROC = 0.76; p = 0.0071). ML and GEP were modestly correlated (r = 0.28), consistent with associations between ML and GEP in Moffitt (r = 0.11) and TCGA databases (r = 0.29). When jointly modeled, ML was significantly associated with response (p = 0.0078) after adjusting for GEP (also significant; p = 0.0251). Data from additional patients will also be presented. Conclusions: These data show that ML and GEP are independently predictive biomarkers of response to pembrolizumab in multiple tumor types, suggesting that tumor antigenicity and T cell infiltration may provide complementary information for expected pembrolizumab activity. As measures of distinct, yet common features of a PD-1 checkpoint blockade responsive tumor, ML and GEP may have utility in characterizing responses to anti PD-1 and other cancer immunotherapies. Clinical trial information: NCT01848834; NCT02054806.
Trastuzumab versus MYL-1401O (a proposed trastuzumab biosimilar) in a phase I bioequivalence study: In vivo and in vitro immunomodulation.
Authors: Audran RÃÂ©gine , Chtioui Haithem , Thierry Anne-Christine , Mayor Carole , Vallotton Laure , Dao Kim , Rothuizen Laura , Maghraoui Ali , Pennella Eduardo J. , Brunner-Ferber Françoise , Buclin Thierry , Spertini Francois
Published: November 2019

10

Background: Trastuzumab is a humanized monoclonal antibody targeting breast cancer cells overexpressing the HER2-oncoprotein. During a Phase-I single centre, single dose, randomized, double-blind, cross-over study assessing the bioequivalence of a proposed trastuzumab biosimilar (MYL-1401O) versus the initially marketed drug (Herceptin), we investigated in addition a large panel of pharmacodynamics parameters comparing the immunomodulatory activity of both drugs. Methods: 22 healthy males were included, 19 subjects receiving randomly a single intravenous infusion of MYL-1401O and 22 of Herceptin, separated by 16 to 22 week wash-out. Blood samples drawn pre- and post- infusion were assessed for in vivo serum cytokines induction (IL-1β, IL-2, IL-6, IL-10, IL-12, TNF-α, GM-CSF and IFN-γ) whereas the impact of treatment on mononuclear cell subsets and their level of activation was tested ex vivo. Volunteers’ PBMC (peripheral blood monocnuclear cells) were stimulated in vitro with recall antigens and mitogen for cytokine production. At baseline, we performed in addition a cytokine release assay on PBMC upon stimulation with trastuzumab as a preclinical safety test. Results: Trastuzumab infusion induced a transient and weak peak of serum IL-6 at 6h, and a modulation of mononuclear cell subset profile and level of activation. Notably CD16+ cells frequency decreased at 3h and peaked at 48h. Except for CD8+ T cells, there were no significant differences between Herceptin and its proposed biosimilar ex vivo. PBMC stimulated in vitro with trastuzumab secreted IL-6, TNF-a, IL-1β, GM-CSF, IFN-γ, and IL-10, but no IL-2. There was no significant difference between the two mAbs. Conclusions: Based on these in vivo, ex vivo and in vitro experiments, there is a strong assumption that MYL-1401O is biosimilar to the reference drug Herceptin for its immunomodulation properties as already proven for its bioequivalence. Clinical trial information: 2011-001406-94.
Activity of CBT-501, a novel humanized anti-PD-1 antibody, in a humanized genetically engineered mouse model of colon carcinoma.
Authors: Zhou Qing , Nian Weihong , Sun Ziyong , Qiu Junzhuan , Wang Zhun , Choy Gavin , Redkar Sanjeev
Published: November 2019

108

Background: CBT-501 (Genolimzumab, GB-226) is a novel humanized mAb (IgG4k) against human Programmed Death-1 (PD-1). CBT-501 demonstrated highly specific binding to PD-1 of human (KD= 505 pM) and cynomolgus (KD= 7.2 nM). It did not completely block the binding of nivolumab or pembrolizumab, suggesting a novel epitope. CBT-501 efficiently inhibited the binding of PD-L1/L2 to PD-1 through a competitive action for both human and monkey. It enhanced human T-cell activation in the Mix Lymphocyte Reaction (MLR), as shown by increased release of IL-2 and INF-γ. The Fc-fragment bound to FcRn dose-dependently without any effector function. The PK/PD studies in cynomolgus showed a linear dose exposure, with a T1/2of 115-142 h for single and repeat dosing. The PD-1 receptor occupancy after a single dose was dose-dependent reaching 100% at peak, maintained after multiple dosing and continued for 6 wk after last dose. Single and repeat dose toxicology studies in cynomolgus showed weight gain of the thyroid and mild expansion of the thyroid follicles at the highest dose of 100 mg/kg which were reversible. No abnormal drug-related toxicity was found. In rhesus vaccinated with adenovirus-mediated Simian Immunodeficiency Virus, biweekly IV CBT-501 promoted the antigen-specific T-cell response, as shown by ELISPOT and intracellular staining of IL-2, INF-γ and TNF-α in PBMC and T-cell subpopulation. Due to lack of immunoselectivity to mouse PD-1, investigation of the in vivo anti-tumor activity in rodent model is challenging. Methods: Crown Bioscience has developed humanized genetically engineered mouse model expressing human PD-1 extracellular domain with a fully functional murine immune system. Subcutaneous colon adenocarcinoma MC38 syngeneic tumor draft was established in mice. Results: CBT-501 injected IP q2w×3wk demonstrated significant anti-tumor activity. The tumor growth reduction was dose dependent with comparable or improved activity over nivolumab, and with the highest inhibition rate at 83%. Conclusions: Based on the strong nonclinical data and differentiation, and an acceptable safety profile, a first in human Phase 1 trial is planned in the Asia-Pacific region.
Analysis of the newly defined immune checkpoints in pancreatic cancer.
Authors: Wada Satoshi
Published: November 2019

109

Background: Pancreatic cancer has not been improved with the five-year survival rate for more than 10 years, which is a refractory cancer of poor prognosis. Development of novel therapeutic agents has been desired, because the standard chemotherapy has not been satisfied yet. Anti-tumor immunotherapy is expected for less burden on the patient in recent years. However, it is limited by multiple tolerogenic mechanisms related to the tumor, the host immune system, and the tumor microenvironment. Therefore satisfactory results are not obtained in the clinical trials for several vaccinations. In this experiment, we focused on negative co-signaling molecules, especially immune checkpoints. The purpose of the present study was to investigate the relationship between the expression of immune checkpoints and clinico-pathological findings, including prognosis in pancreatic cancer patients. Methods: We enrolled 81 patients (52 men and 29 women; age range: 36-84 years) with pancreatic cancer who underwent curative resection with lymphadenectomy at Gunma University Hospital and Saiseikai Maebashi Hospital (Maebashi, Japan) between 2000 and 2011. Patients who had undergone palliative resection, preoperative chemotherapy and radiation therapy were excluded from this study. Furthermore, none of the patients enrolled in this study had synchronous or metachronous cancer in other organs. Results: Immunohistochemical staining of immune checkpoints in primary pancreatic tissue was performed. Patients with high B7-H4 expression had a significantly poorer prognosis than those with low B7-H4 expression. It was also seen in B7-H3 expression in this experiment. Furthermore, in analysis for the expression of CD8, IFN-γ and FOXP3, patients with high FOXP3 expression had a significantly poorer prognosis than those with low FOXP3 expression. Conclusions: The B7-h3 and B7-H4 are negative co-stimulatory molecules and a novel prognostic marker related to the T-cell mediated immune response. It’s pathway may be a molecular target for controlling tumor progression in patients with pancreatic cancer.
Long-term follow-up of DNA engineered bi-shRNA furin GMCSF plasmid/autologous tumor induced immune response in patients with advanced solid tumors (phase I trial).
Authors: Nemunaitis John J. , Barve Minal A. , Melnyk Anton , Wallraven Gladice , Manning Luisa , Senzer Neil N.
Published: November 2019

110

Background: Previously we described safety and evidence of activity to VigilÒ (FANGÒ) and identified relationship of survival advantage to ELISPOT “+” Vigil treated patients (Senzer N et al. Long Term Follow Up: Phase I Trial of “bi-shRNA furin/GMCSF DNA/Autologous Tumor Cell” Immunotherapy (FANG) in Advanced Cancer. J Vaccines Vaccine 2013; 4: 209. doi: 10.4172/2157-7560.1000209). We have updated survival and long-term ELISPOT assessment of this same group of patients. Methods: A non-randomized phase 1 trial of patient treated with Vigil immunotherapy (n=39) compared to a similar matched comparator (MC) group not treated with Vigil (n=35). Results: Trial results suggest survival benefit without evidence of Vigil related toxicity (no ≥ grade 3). γ-IFN ELISPOT served as a biomarker for response and shows correlation with survival. 22/30 evaluable Vigil-treated patients showed ELISPOT conversion (73%) from negative to positive (threshold of ≥10 spots from baseline) during treatment. Durable evidence of γIFN-secreting circulating cytotoxic T cells was observed (up to 30months from treatment start), suggesting induction of memory T effector cells. Survival correlation demonstrated significant benefit of ELISPOT “+” Vigil treated patients (median OS of 784 days, 26.1 months) compared to ELISPOT “-“ patients (median OS of 353 days, 11.77 moths), and supports immune-related survival benefit compared to MC (median OS of 122 days, 4 months), (Table). Conclusions: Vigil immunotherapy induces a significant immune response. Detailed analysis of the immune effector population phenotype is underway to determine optimal correlation with survival. Clinical trial information: NCT01061840. [Table: see text]
Effects of TTI-621 (SIRPαFc) on CD47 and serum cytokines associated with phagocytosis in subjects with relapsed, refractory hematologic malignancies: Pharmacodynamic findings from a first-in-human clinical trial.
Authors: Johnson Lisa , Pillai Raju K. , King Rebecca L. , Ansell Stephen Maxted , Chen Robert W. , Flinn Ian , Maris Michael B. , Irwin Meghan , Sievers Eric L. , Petrova Penka S. , Uger Robert A.
Published: November 2019

112

Background: CD47 delivers an anti-phagocytic (“do not eat”) signal by binding SIRPα on macrophages and is overexpressed on many cancers enabling immune surveillance evasion. TTI-621 is a decoy receptor that promotes phagocytosis of tumors by blocking CD47 and engaging activating FcR on macrophages. A first-in-human, phase 1a, open label, multicenter study (NCT02663518) evaluating safety and tolerability of weekly, intravenous infusions of TTI-621 was conducted. As of October 12, 2016, 18 subjects were treated with TTI-621 with doses of 0.05-0.3 mg/kg. Multiple pharmacodynamic and hematologic assessments were performed to characterize the effects of TTI-621 and better understand the mechanism of action. Methods: Serial blood draws were analyzed for CD47 receptor occupancy (RO), complete blood counts, cytokines, T cell repertoire changes and CD47, SIRPα and FcR sequences. CD47 expression, macrophage and T cell infiltration was assessed on biopsies by IHC. Results: Peripheral CD47 levels varied considerably between subjects and were inversely correlated with RO. A maximum of 80% RO was achieved on peripheral leukocytes. Circulating monocytes, neutrophils, and lymphocytes were transiently decreased at the end of the TTI-621 infusion and generally returned to baseline levels by 24 to 72 hours. Platelet counts decreased immediately after TTI-621 infusions, typically recovering to baseline levels prior to the subsequent treatment cycle. Hemoglobin levels did not meaningfully change during treatment. Cytokines associated with macrophage activation or phagocytosis, notably MIP-1α, MIP-1β, TNF-α and IP-10 were elevated in a dose dependent manner. Conclusions: Systemic administration of TTI-621 leads to CD47 blockade and dose dependent increases in cytokines associated with phagocytosis, temporally associated with reversible thrombocytopenia, suggesting enhanced macrophage mediated clearance of circulating platelets followed by a robust marrow regenerative response. Enrollment into multiple expansion cohorts addressing a wide range of hematopoietic cancers is ongoing. Clinical trial information: NCT02663518.
Conversion of CCL18-recruited naïve CD4+ T cells to tumor-infiltrating regulatory T cells in breast cancer and suppression of antitumor immunity.
Authors: Su Shicheng , Liao Jianyou , Liu Jiang , Liu Qiang , Song Erwei
Published: November 2019

114

Background: Tumor-infiltrating regulatory T cells (Tregs) play a central role in tumor immunosuppression. However, it remains unclear whether they are directly recruited from peripheral blood or converted from infiltrating naive T cells. Methods: We use full-length TCR ¦Á/¦Â repertoire to analyse the difference of T cell subsets from peripheral blood, primary tumors and draining lymph nodes in patents. Results: Infiltration of naive CD4+ T cells and Tregs are closely correlated, both indicating poor prognosis for breast cancer patients. Naive CD4+ T cells in the tumors are recruited by tumor-associated macrophages (TAMs) via CCL18. In addition, naive T cells and memory T cells exhibit distinctive chemotactic response due to different expression of regulator of G-protein signaling 1(RGS1). Specific silencing CCL18 receptor-PITPNM3 in naive CD4+ T cells using CD4 aptamer-siRNA blocks their chemotaxis, and thus reduces infiltrating Tregs and inhibits tumor progression in humanized mice. By comparison, silencing RGS1 in memory CD8+ T cells using CD8 aptamer-siRNA enhance their recruitment to tumors and anti-tumor immune response in vivo. Conclusions: These findings provide mechanistic insights for Treg enrichment in breast cancer and suggest that modification of the CCL18-PITPNM3-RGS1 signaling pathway may be an attractive strategy for anticancer immunotherapy.
Toll like receptors as a risk factor for early death in elderly cancer patients.
Authors: Telles O Lima Jurema , Mello Maria Julia Gonçalves , Thuler Luiz Claudio Santos , Telles Sales Letícia , Santaliz de Godoy Moreno Marina , Bergmann Anke , Jatahy Kleber , Resende Lira Clara Cinthia , Bezerra Cintra Evandro , Delgado Soriano Gabriela , Torres Leuridan
Published: November 2019

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Background: the elderly cancer group is a heterogeneous and growing one. In the elderly, the genes of toll-like receptors (TLR) have been described as related to the immunosenescence process and carcinogenesis. The relationship of this gene family with carcinogenesis and immunoregulatory responses seems a promising field. Methods: Between 2015 and 2016, a prospective cohort study in 445 elderly patients with incident cancer ( ≥ 60 years) at the time of admission, assessed and collected sociodemographic and clinical variables and collected analysis of peripheral blood in translational exploratory study. Determination of TLR2 and TLR9 was performed by flow cytometry with monoclonal antibodies anti-TLR2 and TLR9 in the peripheral blood at the beginning of anti-cancer therapy in older patients with cancer. Results: 445 elderly patients were included with incident cancer, age of 71.13 years (SD 7.41) means. Most were male (50.9%) and self-reported black or brown skin color (71.4%). The topography of prostate cancer was more frequent (29.4%), followed by the digestive system (24.3%). They were mostly patients with advanced stage (III and IV) at nutritional risk (52.6%) and reported using up to 5 medications (77.2%). Patients were followed for an average of 128 days (SD 54.37). During this period, there were 62 deaths (13.9 %) with a mean overall survival of 165 days (95% CI 161.25 to 170.02). There are significant differences in the percentage values of TLR9 (p = 0.0009) and TLR2 (p = 0.04) in monocytes between the groups of patients who died in less than 6 months of treatment and those who survived. Conclusions: Overexpression of TLR9 and TLR2 may be a protective factor for the occurrence of early death in elderly patients with cancer.
Plinabulin as a novel small molecule clinical stage immuno-oncology agent for NSCLC.
Authors: Mohanlal Ramon W. , LLoyd Ken , Huang Lan , Bazhenova Lyudmila
Published: November 2019

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Background: Plinabulin (Plin) is a small molecule with tumor-inhibiting and immune-enhancing effects. In preclinical studies, Plin induces dendritic cell maturation and related IL-1β, IL-6 and IL-12 release, enhances antigen-specific CD4 T-cell proliferation, reduces regulatory T-cells and M2 macrophages in tumor tissue. Plin had synergistic efficacy with checkpoint inhibitors (PD-1 +/- CTLA-4) in animal models of tumor growth. Plin is targeting critical steps in the immune-cascade, independent from that of PD-1/PD-L1 binding. Methods: In a Phase 2 clinical trial, patients (pts) with and without measurable lung lesion were randomized to docetaxel (D) alone at 75 mg/m2 or Plin with D (Plin+D), in 30 mg/m2 or 20 mg/m2 Plin cohorts, given every 3 weeks (NCT00630110). The primary efficacy endpoint was mOS. Secondary endpoints were safety assessments, DOR, PFS, ORR. PD-L1 tumor status was not characterized. Results: In the 30 mg/m2 cohort, n=50 patients received 30 mg/m2 Plin+ D and n=55 patients received D alone; 72% had measurable lung lesion. In pts with a measurable lung tumor, Plin+D (n=38) was numerically more effective vs. D alone (n=38). mOS, PFS and ORR were 11.3 mo, 3.7 mo, and 18% respectively for Plin+ Doc, and 6.7 mo, 2.9 mo, and 10.5% resp for D alone. DOR (a marker of immune effect) with the Plin+D vs. D alone was 12.7 mo vs. 1 mo (p<0.05). Plin+D did not induce immune-related adverse events (IR-AEs), but prevented D- induced neutropenia (p<0.0002), and D-dose-reduction due to toxicity. The 30 mg/m2 Plin cohort was more effective than the 20 mg/m2 cohort as an anticancer agent. Conclusions: Plin added to D increases efficacy vs. D alone most likely through an immune-related mechanism independent from PD1/PD-L1 intervention and mitigates the known D-related safety concerns, without increasing IR-AEs. The Plin+D combination might represent an effective and cost-effective alternative to Nivolumab, in particular in PD-L1 negative NSCLC. A global phase 3 trial with Plin+D vs. D alone is underway in pts with at least one measurable NSCLC lesion. Clinical trial information: NCT00630110.
Pan-cancer opportunities for off-label immunotherapy based on nonsynonymous mutation burden.
Authors: Knepper Todd Cory , Teer Jamie K. , Walko Christine Marie , McLeod Howard L.
Published: November 2019

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Background: In an effort to better predict which cancer patients (pts) are more likely to derive clinical benefit from immunotherapy, an association between increased non-synonymous mutation burden and likelihood of clinical response has been described. This study reports the mutation burden of pts with a variety of advanced cancers within a real-world clinical genomics database in order to highlight opportunities where off-label immunotherapy can be considered. Methods: For each solid cancer assayed by FoundationOne within Moffitt’s Clinical Genomic Action Committee (CGAC) database, a non-synonymous mutation burden was determined. Pts were then grouped by cancer type and descriptive statistics used to describe the mutation burden. For cancers with at least 5 cases, the percentage of pts with a non-synonymous mutation burden exceeding a threshold shown to be suggestive of clinical benefit from immunotherapy ( ≥ 13) is reported. The threshold was derived via linear regression from the “intermediate-high” mutation burden classification of the FoundationOne assay. Results: 520 pts received a FoundationOne test. The median non-synonymous mutation burden was 7 and the mean was 11.4. More pts with cancers that have demonstrated response to immunotherapy had a mutation burden exceeding the threshold (19.7% for lung – 45.0% for melanoma). Pt tumors from cancer types without immunotherapy indications sometimes (5.0% for salivary gland – 10.5% for breast and colorectal cancer) exceeded mutation threshold suggestive of benefit. Conclusions: A meaningful proportion of pts with cancer types without FDA-approved immunotherapies have a mutation burden for which immunotherapy may represent a clinical consideration. [Table: see text]
Expansion of tumor infiltrating lymphocytes (TIL) from bladder cancer.
Authors: Poch Michael Adam , Hall MacLean S , Kodumudi Krithika Nandakumar , Croft Cortlin , Fishman Mayer N. , Mullinax John , Mule' James J , Sarnaik Amod , Pilon-Thomas Shari
Published: November 2019

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Background: Patients with advanced bladder cancer have limited therapeutic options resulting in a median overall survival (OS) between 12 and 15 months. At our institution, adoptive cell therapy (ACT) using tumor infiltrating lymphocytes (TIL) has resulted in a durable median OS of 52 months in patients with metastatic melanoma. Immune-mediated anti-tumor responses have been previously shown in bladder cancer, therefore we investigated the phenotype and function of TIL expanded from bladder tumors to establish feasibility of ACT for the treatment of bladder cancer. Methods: Tumor specimens, including primary bladder tumors and lymph node metastases, were collected from 29 bladder cancer patients having standard of care tumor resection, who also had consented to an IRB-approved protocol for TIL generation. The tissue was minced into fragments, placed in individual wells of a 24-well plate, and propagated in high dose IL-2 for four weeks. TIL were considered expanded if they propagated to fill ≥2 wells. The remaining tumor material was digested into a single cell suspension and frozen. TIL were phenotyped by flow cytometry and assessed for autologous tumor reactivity through co-culture with tumor digest and IFN-gamma ELISA. Results: Transitional cell bladder tumors were cultured from 23 patients, of whom 19 (83%) demonstrated TIL expansion. Microbial contamination precluded TIL growth in six specimens. TIL were cultured from 9/12 (75%) patients with preceding chemotherapy and 10/11 (91%) who were chemotherapy naive. Expanded TIL were predominantly CD3+(median 63%, range 10-87%) with a median of 30% CD8+ T cells (range 5-70%). Eight of 15 tested samples (53%) contained TIL that secreted IFN-gamma in response to autologous tumor. Conclusions: The study establishes the practical first step towards an autologous TIL therapy process for therapeutic testing in patients with bladder cancer. Human bladder cancer tissue can be used to expand tumor-specific TIL in vitro. TIL were also expanded from patients that received chemotherapy prior to tumor resection. Future efforts will explore the ability to further expand bladder TIL cultures to clinically meaningful numbers to develop novel ACT strategies for patients with this diagnosis.
CD70 as a novel target of CAR-T-cell therapy for gliomas.
Authors: Jin Linchun , Ge Haitao , Yang Changlin , Long Yu , Chang Yifan(Emily) , Mu Luyan , Sayour Elias , De leon Gabriel , Wang Qiong J. , Yang James C. , Drake Jeffrey , Kubilis Paul , Lu Dunyue , Kong Yingjun , Hu Li , Jiping Qi , Gu Yunhe , Lin Zhiguo , Mitchell Duane Anthony , Huang Jianping
Published: November 2019

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Background: Gliomas are the most common primary malignant brain tumor and are uniformly lethal. Cancer immunotherapy has the potential to target gliomas; however, its antitumor effects are restricted by limitations in clinically useful tumor specific targets. Chimeric antigen receptor modified T-cell (CAR-T) therapy is a highly promising option for cancer treatment, due to its combination of precision antibody recognition and T-cell tumor-specific killing. CD70 is an antigen expressed by limited subsets of normal lymphocytes and dendritic cells but is aberrantly overexpressed by glioma cells, which makes it an outstanding glioma-antigen target. Methods: The gene and protein expression of CD70 were evaluated to identify its potential as a glioma target. Human and mouse versions of CD70-specific CAR-T cells were generated, and human primary GBM lines as well as murine lines (GL-261, KR-158B) were used as human and mouse tumor targets, respectively. The antitumor effect of the human and mouse CD70-sepecific CARs were tested in vitro and in orthotopic xenograft and syngeneic murine models. Results: CD70 is only overexpressed by tumor cells in a subset of low-grade gliomas and GBM. The elevated gene and protein expression are associated with increased tumor grade and poor patient survival. Co-culturing CD70-specific CAR-T cells with CD70-positive glioma cells resulted in potent secretion of IFN-gamma and tumor-specific killing in a CD70-dependent manner. Irradiation enhances CD70 expression on glioma cells and thus increases CAR T-cell recognition. Adoptive transfer of the human and mouse CD70 CAR-T cells resulted in tumor regression of immunocompetent and immunodeficient mice, respectively. Conclusions: CD70 can be an excellent tumor target for gliomas, and CD70-specific CAR-T cells have potent antitumor activity against CD70-positive gliomas both in vitro and in vivo. Our study provides crucial preclinical evidence to support the future clinical application of CD70 CAR-T cells to treat gliomas.