bluuebunny/crossref_metadata_2025 huggingface dataset:
Potential impact of climate change on plant invasion in the Republic of Korea
Authors: Adhikari Pradeep , Jeon Ja-Young , Kim Hyun Woo , Shin Man-Seok , Adhikari Prabhat , Seo Changwan
Published: November 2019

Background

Invasive plant species are considered a major threat to biodiversity, ecosystem functioning, and human wellbeing worldwide. Climatically suitable ranges for invasive plant species are expected to expand due to future climate change. The identification of current invasions and potential range expansion of invasive plant species is required to plan for the management of these species. Here, we predicted climatically suitable habitats for 11 invasive plant species and calculated the potential species richness and their range expansions in different provinces of the Republic of Korea (ROK) under current and future climate change scenarios (RCP 4.5 and RCP 8.5) using the maximum entropy (MaxEnt) modeling approach.

Results

Based on the model predictions, areas of climatically suitable habitats for 90.9% of the invasive plant species are expected to retain current ecological niches and expand to include additional climatically suitable areas under future climate change scenarios. Species richness is predicted to be relatively high in the provinces of the western and southern regions (e.g., Jeollanam, Jeollabuk, and Chungcheongnam) under current climatic conditions. However, under future climates, richness in the provinces of the northern, eastern, and southeastern regions (e.g., Seoul, Incheon, Gyeonggi, Gyeongsangnam, Degue, Busan, and Ulsan) is estimated to increase up to 292%, 390.75%, and 468.06% by 2030, 2050, and 2080, respectively, compared with the current richness.

Conclusions

Our study revealed that the rates of introduction and dispersion of invasive plant species from the western and southern coasts are relatively high and are expanding across the ROK through different modes of dispersion. The negative impacts on biodiversity, ecosystem dynamics, and economy caused by invasive plant species will be high if preventive and eradication measures are not employed immediately. Thus, this study will be helpful to policymakers for the management of invasive plant species and the conservation of biodiversity.
The expansion and validation of a new upper extremity item bank for the Patient-Reported Outcomes Measurement Information System® (PROMIS)
Authors: Kaat Aaron J. , Buckenmaier Chester “ Trip” , Cook Karon F. , Rothrock Nan E. , Schalet Benjamin D. , Gershon Richard C. , Vrahas Mark S.
Published: November 2019

Background

The Patient-Reported Outcomes Measurement Information System® (PROMIS) includes a Physical Function (PF) item bank and an Upper Extremity (UE) item bank, which is composed of a subset of items from the PF bank. The UE item bank has few items and known ceiling effects. Therefore, this study aimed to expand the item bank to assess a wider range of functioning. With the additional content, other psychometric properties—improved content validity, item bank depth, range of measurement, and score reliability—were also evaluated. We convened an expert panel to review potential items, and then conducted psychometric analyses on both extant and newly-collected data.

Results

Expert focus groups reviewed the PF item bank for items that were “sufficiently” related to upper extremity functioning for inclusion in the expanded UE item bank. The candidate item bank was quantitatively evaluated in a new sample of 600 people. The final items were calibrated in an aggregated dataset (n = 11,635) from two existing datasets, and the newly collected sample. The original UE item bank included 15 items. After expert review and quantitative evaluation, 31 items were added. The combined 46 items were calibrated using item response theory (IRT). Then computer adaptive tests (CATs) were simulated based off of the psychometric results. These indicated that the new UE item bank has an extended measurement range compared to the original version.

Conclusions

The expanded PROMIS UE item bank assesses a wider range of upper extremity functioning compared to the initial UE item bank. However, ceiling effects remain a concern for unimpaired groups. The new UE item bank is recommended for individuals with known or suspected upper extremity limitations.
Identification of proteins differentially expressed by glutamate treatment in cerebral cortex of neonatal rats
Authors: Kang Ju-Bin , Park Dong-Ju , Koh Phil-Ok
Published: November 2019

Glutamate leads to neuronal cell damage by generating neurotoxicity during brain development. The objective of this study is to identify proteins that differently expressed by glutamate treatment in neonatal cerebral cortex. Sprague-Dawley rat pups (post-natal day 7) were intraperitoneally injected with vehicle or glutamate (10 mg/kg). Brain tissues were isolated 4 h after drug treatment and fixed for morphological study. Moreover, cerebral cortices were collected for protein study. Two-dimensional gel electrophoresis and mass spectrometry were carried out to identify specific proteins. We observed severe histopathological changes in glutamate-exposed cerebral cortex. We identified various proteins that differentially expressed by glutamate exposure. Identified proteins were thioredoxin, peroxiredoxin 5, ubiquitin carboxy-terminal hydrolase L1, proteasome subunit alpha proteins, isocitrate dehydrogenase, and heat shock protein 60. Heat shock protein 60 was increased in glutamate exposed condition. However, other proteins were decreased in glutamate-treated animals. These proteins are related to anti-oxidant, protein degradation, metabolism, signal transduction, and anti-apoptotic function. Thus, our findings can suggest that glutamate leads to neonatal cerebral cortex damage by regulation of specific proteins that mediated with various functions.
Multiple b values of diffusion-weighted magnetic resonance imaging in evaluation of solid head and neck masses
Authors: Abou khadrah Rania Sobhy , Imam Haytham Haroon
Published: November 2019

Background

Differentiation between malignant and benign masses is essential for treatment planning and helps in improving the prognosis of malignant tumors; the aim of this work is to determine the role of diffusion-weighted magnetic resonance imaging (DW-MRI) and the apparent diffusion coefficient (ADC) in the differentiation between benign and malignant solid head and neck masses by comparing diagnostic performance of low b values (0.50 and 400 s/mm2) versus high b values (800 and 1000 s/mm2) and comparing the result with histopathological finding.

Results

The study included 60 patients (34 male and 26 female) with solid head and neck masses > 1 cm who referred to radiodiagnosis department for MRI evaluation. Multiple b values were used 50, 400, 800, and 1000 s/mm2 (at least 2 b values). DWI and ADC value of all 60 patients were acquired. Mean ADC values of both malignant and benign masses were statistically measured and compared, and cut off value was determined. Solid head and neck masses in our study DWI with the use of high b value 800 and 1000 s/mm2 were of higher significance (P value 0.001*). There was a significant difference in the mean ADC value between benign and malignant masses (P < 0.01); solid masses were divided into 2 categories: (a) malignant lesions 46.7% (n = 28) with mean ADC value (0.82 ± 0.19) × 10−3 s/mm2 and (b) benign lesions 53.3% (n = 32) with mean ADC value (2.05 ± 0.46) × 10−3 s/mm2) with ADC cutoff value of 1.0 × 10−3 s/mm2 and 94% sensitivity, 93% specificity, negative predictive value (NPV) = 94%, positive predictive value (PPV) 93%, and an accuracy of 93.5%.

Conclusion

The DWI with ADC mapping were valuable as non-invasive tools in differentiating between benign and malignant solid head and neck masses. The use of high b value 800 and 1000 s/mm2 was of higher significance (P value 0.001*) in differentiation between benign and malignant lesion than that with low b values 0, 50, and 400 s/mm2 (0.01). The mean ADC values were significantly lower in malignant solid masses. Attention had to be paid to the choice of b values in MRI-DWI in the head and neck region.
MRI comparative study of levator ani muscle changes in nulliparous and multiparous females
Authors: Azzam Heba , Halim Manal , El-Assaly Hany , Heiba Aya
Published: November 2019

Background

Pelvic floor dysfunction is known to be among the principal factors influencing public health, regarding frequency, cost and effect on women’s quality of life. Radiographic assessment of the pelvic floor function and anatomy plays a vital role in the recognition of pelvic floor defects. The aim of this study is to detect the postpartum-related levator ani muscle changes thus defining the relationship between the vaginal deliveries and the etiology of pelvic floor dysfunction in order to provide guidelines to decrease the incidence of pelvic floor injuries during parturition and guide the treatment plan.

Results

There was a significant difference in the puborectalis muscle thickness between the case and control groups in the right puborectalis (P value ≤ 0.001) and in the left puborectalis (P value (≤ 0.001) as well as significant midpoint thickness (P value = 0.03) with 46.2% puborectalis muscle injury in the case group compared with none in the control group.

Conclusion

Pelvic floor MRI is highly recommended as it is a contrast-free modality that allows for both anatomical and functional analysis. Its incorporation in the routine postpartum assessment will allow early detection of abnormalities even in asymptomatic cases thus ensuring proper management and preventing the development of pelvic floor dysfunction predisposed to by repeated vaginal deliveries.
Chemical and pharmacological evaluation of manglicolous lichen Roccella montagnei Bel em. D. D. Awasthi
Authors: Tatipamula Vinay Bharadwaj , Vedula Girija Sastry , Sastry A. V. S.
Published: November 2019

Background

Lichen is a composite organism composed of fungus in association with algae or cyanobacteria. Particularly, lichen betide to mangroves are named as manglicolous lichens. From the folklore, lichen extracts were used in the management of many infections and diseases.

Results

The chemical investigation of acetone extract of manglicolous lichen Roccella montagnei (RM-Ac) yielded nine known metabolites namely divarinolmonomethylether (1), ethyl divaricatinate (2), divarinol (3), orcinol (4), methyl 2,6-dihydroxy-4-methylbenzoate (5), haematommic acid (6), atranol (7), ethyl haematommate (8) and ethyl orsellinate (9). Except 4, all are for the first reported from this species. The RM-Ac and its metabolites (1–9) were screened for antioxidant (DPPH, ABTS and superoxide free radical assays), anti-inflammatory, anticancer (SRB assay using A549, DLD-1, FADU, HeLa and MCF-7) and acute toxicity studies. The pharmacological results showed that compounds 6 and 8 depicted potent inhibitory profile against ABTS free radical (with IC50 value of 40.0 and 40.5 μg/mL, respectively) and protein denaturation (with IC50 value of 435 and 403 μg/mL, respectively). LD50 of RM-Ac was found to be above 2 g/Kg body weight. Moreover, the RM-Ac showed prominent inhibition of formalin-hind albino rat paw oedema at both the tested doses, i.e., 100 and 200 mg/Kg b.w than that of the standard drug (indomethacin). Furthermore, the compounds 6 and 8 exhibited significant degree of specificity against HeLa, FADU and A549; besides, they showed very little degree of specificity against NHME cell line specifying less toxicity to normal cells.

Conclusion

It can be concluded that the manglicolous lichen R. montagnei has an aptitude to act against free radicals, inflammation and cancer, and the main metabolites responsible for its biological activity are 6 and 8.
Comprehensive genomic analysis with immunological parameters to obtain the biomarkers for immune checkpoint inhibitors (ICIs) in non-small cell lung cancer (NSCLC).
Authors: Owada Yuki , Muto Satoshi , Takagi Hironori , Inoue Takuya , Watanabe Yuzuru , Yamaura Takumi , Fukuhara Mitsuro , Okabe Naoyuki , Matsumura Yuki , Hasegawa Takeo , Tanaka Daisuke , Ito Emi , Nanamiya Hideaki , Imai Jun-ichi , Isogai Takao , Watanabe Shinya , Suzuki Hiroyuki
Published: November 2019

107

Background: Several kinds of predictors of ICIs efficacy in NSCLC have been analyzed. PD-L1 expression of tumor, frequency of tumor infiltrating lymphocytes (TIL) and microsatellite instability are cited as potential predictors of treatment efficacy of ICIs, but these still seem to be immature. Lately, mutation load in tumor, presumably involving to neo-antigens in tumor tissue, is also thought to be one of more probable predictors for ICIs. However, the hypothesis is still limited and it is very hard to analyze the mutation load routinely in clinic. We here describe more definitive and easily detecting factors by using Next generation sequencing (NGS) and by analyzing conventional immunological and molecular parameters. Methods: 94 patients with NSCLC who undergone resection in our institution were subjected for this study. Mutation load in tumor samples was analyzed by whole exome sequencing using NGS. The major gene alteration was also analyzed by NGS. TILs and PD-L1 expression in tumor were immunohistochemically assessed by using CD8 and SP142 (anti-PD-L1 antibody). Results: The median value of mutation load was 54 genes (10-363). The factors; squamous cell carcinoma, EGFR-mutation negative, and TP53 alteration positive indicated close connection with higher mutation load. PD-L1 expression, TILs, gender, age, and etc. were not associated with mutation load. Multiple regression analysis shows that EGFR-mutation negative and TP53 alteration positive contribute to higher mutation load (p = 0.018, p = 0.015). Conclusions: Mutation load in tumor could be associated with EGFR-mutation and TP53 alteration status. These profiles could potentially be more simple biomarkers in treating ICIs.
Tumor associated macrophages antagonize antitumor effect of chemotherapy.
Authors: Chen Fei , Su Shicheng , Song Erwei
Published: November 2019

120

Background: Tumor associated macrophages play an essential role in tumor progression, promoting angiogenesis, immunosupression, invasion and metastasis. However, TAMs regulation of tumor responses to chemotherapy is complicated. In different type of treatment or tumor model, TAMs can either enhance or antagonize antitumor effect of chemotherapy. Methods: We isolated TAMs from breast cancer tissues, or PBMC stimulated with culture medium of mesenchymal breast cancer cells and cocultured with breast cancer cells MCF-7 for 6-10days. After cocultured with TAMs, we treated MCF-7 with docetaxel. In vivo, we use MCF-7 with or without TAMs to generate xenografts in NOD/SCID mice and treated with docetaxel. To testify if the antagonize was dependent on specific treatment, we also treated breast cancer cells with ciaplatin. Results: With coculture of TAMs, we obversed ruduced apoptosis of docetaxel treated breast cancer cells. In mice model, TAMs co-injection xenografts antagonized the antitumor effect of docetaxel.Consisted with previous results, TAMs coculture antagonize the antitumor effect of cisplatin, suggesting the antagonize of TAMs was independent on cell cycle. Conclusions: Collectively, our study indicated the antagonize of TAMs towards chemotherapy.
GRM1 to induce M-CSF and CCL2 expression to lead to an upregulation of M2 macrophages in melanoma.
Authors: Li Jiadong , Wen Yu , Jeong Byeong-Seon , Shah Raj , Isola Allison , Silk Ann W. , Chen Suzie , Goydos James Steven
Published: November 2019

122

Background: GRM1 transgenic mice in an immune competent background (C57BL6) develope melanoma with 100% penetrance (TG-3 mice). Ectopic expression of GRM1 in immortalized mouse melanocytes (MelanA) produce cells (MASS20) that are tumorigenic in both SCID and immune competent C57BL6 mice. We have examined the downstream events of GRM1 signaling that allow transformed cells to evade immune destruction. Methods: Cytokine antibody arrays and Western blot were used to examine differences in cytokine expression between MASS20 cells and parental controls. Next, we cultured C57BL6 bone marrow cells in MASS20 conditioned medium for 24 and 48 hours and determined resultant M2 macrophage levels. Finally, we used oral weekly liposome clodronate (LC) treatment to deplete macrophages in murine models to determine the effect on tumor growth. Results: MASS20 cells produce higher levels of M-CSF and CCL2 as compared with control cells. These two cytokines recruit and induce the polarization of tumor associated macrophages (TAM) to increase the M2 fraction resulting in accelerated tumor progression. Culturing C57BL6 bone marrow cells in MASS20 conditioned medium increased expression of the M2 macrophage marker arginase1 at both time points. Weekly oral LC treatment to C57BL6 mice greatly inhibited the growth of MASS20 tumors as compared to growth in control animals. Single-cell suspensions of spleen and tumor cells from these animals were subjected to flow cytometry and showed that LC treatment depleted macrophages by 90% in the spleen, and increased the percentage of cd8+T and cd335+ NK cells in tumors from treated mice as compared to controls. Finally, we treated TG-3 transgenic mice with weekly oral LC and found that treated animals still developed tumors, but progression was greatly slowed, and survival greatly prolonged, compared to untreated animals. Within 1 week of stopping LC in the treated animals there was rapid growth of the tumors. These findings suggest a link between GRM1-signal transduction and TAM-induced suppression of tumor-immunity in melanoma. Conclusions: We conclude that GRM1-induced suppression of tumor immunity occurs through induction of M2 macrophages in the tumor microenvironment.
Effects of in vitro ATRA treatment on human MDSC expansion and function.
Authors: Tobin Richard P. , Jordan Kimberly R , Davis Dana , McCarter Martin
Published: November 2019

125

Background: Myeloid derived suppressor cells (MDSCs) are a heterogeneous population of immature immunosuppressive myeloid cells that are expanded in tumor bearing hosts. Melanoma patients with high frequencies of MDSCs have decreased overall survival and an increased risk of death and disease progression, as well as impaired responses to ipilimumab. Targeting MDSCs to decease their frequency or suppressive activity may provide an effective means to improve the efficacy of anti-cancer therapies. All-trans retinoic acid (ATRA) is a vitamin A derivative currently used to treat APL that may target MDSCs. ATRA differentiates immature myeloid cells into macrophages, dendritic cells, or granulocytes. ATRA has been tested as an MDSC targeting agent in two completed caner clinical trials and is being investigated in several other open and completed trials. Methods: We purified myeloid cells from LRS chambers collected from normal donors. The purified cells were treated with GM-CSF+IL-6, GM-CSF+IL-4, or melanoma conditioned media with or without ATRA. After 5 days of incubation, PCR was used to determine the expression of immunoregulatory genes. Additionally, we used a MLR to determine the effect of ATRA on MDSC’s T cell suppressive function. Results: Here we report that in vitro ATRA treatment decreases the expression of ARG1, NOX1, INOS, and PD-L1. Further, we report that ATRA treatment improved T cell proliferation. Conclusions: Controlling MDSC suppressive function and accumulation may provide a mechanism to improve the efficacy of many current cancer therapies. Here we show that ATRA reduces the expression of the immunosuppressive genes ARG1, NOX1, INOS, and PD-L1 in MDSCs. Additionally, we show that treatment of MDSCs with ATRA decreases MDSC’s ability to suppress T cell proliferation. These results indicate that ATRA may improve the efficacy of conventional and immnunotherapeutic treatments.
Correlation between strength of T-cell response against HPV16 and survival after vaccination with HPV16 long peptides in combination with chemotherapy for late-stage cervical cancer.
Authors: Melief Cornelis Joseph , Gerritsen Winald R. , Welters Marij , Vergote Ignace , Kroep Judith R. , Kenter Gemma , Ottevanger P. B. , Tjalma Wiebren Age , Denys Hannelore , Nijman Hans , van Poelgeest Mariette IE , Reyners An K.L , Velu Thierry J. , Blumenstein Brent A. , Goffin Frederic , Lalisang Roy Iqbal , Stead Richard B. , van der Burg Sjoerd
Published: November 2019

140

Background: Therapeutic vaccination with HPV type 16 synthetic long peptides (HPV16-SLP) results in T cell–mediated regression of HPV16-induced premalignant lesions but fails to install effective immunity in patients with advanced HPV16-positive cervical cancer. We showed that HPV16-SLP vaccination in mice and in patients with advanced cervical cancer patients fosters robust HPV16-specific T cell responses, when combined with chemotherapy. In this study we noted that a single dose of vaccine 2 weeks into the 2nd cycle of chemotherapy was optimal, because at this time the immunosuppressive myeloid cells were down. Methods: We now completed a chemo-immunotherapy study in a larger number of patients with late stage HPV16+ cervical cancer. Three HPV16-SLP vaccine doses were given 2 weeks after the second, third, and fourth cycles of standard chemotherapy. Cohorts of 12 patients each were vaccinated with each of 4 dose levels (20, 40, 100, and 300 µg/ per peptide) of 13 overlapping HPV16 synthetic long peptides (HPV16-SLP) together covering the length of the 2 E6 and E7 proteins. Results: Robust vaccine-induced HPV16-specific T cell responses as assessed by interferon-g Elispot were observed and were sustained throughout the cycles of chemotherapy. These T cell responses were substantially increased in all patients who received HPV16-SLP . In addition, the chemotherapy augmented recall responses to microbial antigens. Such robust T cell responses were not noted in previous trials when similar patients were vaccinated without timing of vaccination during chemotherapy. A marked correlation was observed between the strength of the vaccine-induced immune response and longer-term clinical outcomes such as overall survival. No such correlation exists between the strength of the T cell response against common recall antigens and survival. In addition, a remarkably high proportion of patients survived beyond 20 months after the start of therapy. Conclusions: These results indicate that the survival advantage is specifically related to the strength of the vaccine-induced T cell response and is not due to generally better immuno-competence. Clinical trial information: NCT02128126.
Chimeric antigen receptor (CAR) T cells genetically engineered to deliver IL-12 to the tumor microenvironment in ovarian cancer.
Authors: Yeku Oladapo O. , Purdon Terence , Spriggs David R. , Brentjens Renier J.
Published: November 2019

141

Background: Chimeric antigen receptor (CAR) T cell therapy for solid tumor malignancies has not shown the same degree of clinical efficacy observed in hematologic malignancies such as B-ALL. The presence of an immunosuppressive cellular and cytokine microenvironment has been hypothesized as one reason for the failure of adoptive immunotherapy for solid tumors. In ovarian cancer, myeloid derived suppressor cells (MDSC) and immunosuppressive cytokines in the ascitic microenvironment have been reported. IL-12 is a proinflammatory cytokine produced by macrophages, dendritic cells (DC), and NK cells, and it has been shown to increase proliferation of T cells, induce differentiation of type 1 T helper cells, inhibit regulatory T cells, promote maturation of DCs, and enhance antigen presentation by macrophages. We hypothesize that CAR T cells genetically modified to constitutively secrete IL-12 will overcome a hostile tumor microenvironment in a peritoneal carcinomatosis model of ovarian cancer. Methods: CAR T cells were generated from retroviral transduction of second generation and IL-12 modified CAR’s directed to either an irrelevant CD-19 antigen or Muc16ecto. C57BL/6 mice were inoculated i.p with syngeneic ovarian cancer cells and treated with various CAR T cells. Results: Here we report increased production of IL-12, improved proliferation and cytotoxic activity of 4H1128ζ-IL12 CAR T cells. Further, we show increased levels of inflammatory cytokines at 24 and 48hrs after treatment of tumor-bearing mice, leading to increased survival at advanced stages of disease. Animals treated with 4H1128ζ-IL12 CAR T cells had decreased levels of F4/80+ CD11b+MDSCs. Genetic analysis of recovered MDSCs from the ascites of treated animals showed skewing towards an M1-phenotype via upregulation of cytokines, chemokines, MHC-II, and downregulation of Arg1. Furthermore, clodronate-mediated depletion of TAM’s further enhanced survival in mice treated with 4H1128ζ-IL12 CAR T cells. Conclusions: These results demonstrate the mechanisms of efficacy of localized delivery of IL-12 to the tumor microenvironment by 4H1128ζ-IL12 CAR T cells.
Modulating lncRNA NKILA in tumor-reactive T cells to enhance their trafficking to breast cancer by inhibition of FasL-induced T-cell apoptosis.
Authors: Huang Di , Su Shicheng , Song Erwei
Published: November 2019

143

Background: Adoptive cell therapy (ACT) is a highly personalized cancer therapy that involves administration to the autologous tumor-reactive lymphocytes with direct anticancer activity. In addition to the effective application of ACT for cancer treatment to genetically engineer lymphocytes to express conventional T cell receptors or chimeric antigen receptors, many of the ACT therapy has failed because of the trafficking barrier of T cells. Methods: In vitro, we established tumor-reactive CTL by vaccination with autologous DCs pulsed with MUC1-derived peptides. Moreover, tumor-reactive T cells were transferred to the immunocompromised mice inoculated with autologous patient derived breast cancer xenografts (PDX). Results: In our study, we found that during the induction of tumor-reactive T cells, cytotoxic T lymphocytes (CTL) may undergo massive expansion and have the ability of specific lysis to tumor cells upon appropriate antigenic stimulation via NF-κB pathway activation. However, hyperactive tumor-specific lymphocytes were hypersensitivity to apoptosis signals in tumor milieu, such as Fas ligand. In this research, we showed that NKILA, an NF-κB interacting lncRNA, is up-regulated at the later phase of CTL activation. The up-regulated NKILA suppresses NF-κB activity and sensitizes T cells to apoptosis. In vivo, NKILA knockdown in tumor-reactive T cells upon transferring to the immunocompromised mice inoculated with autologous PDX enhances CTL in xenografts, and thus effectively inhibits tumor growth. Conclusions: Collectively, our findings suggested that NKILA upregulation is responsible for the apoptosis of tumor-responsive CTLs and targeting the lncRNA may provide a novel strategy to enhance the therapeutic efficacy of immune cell transferring for breast cancer.
Phase I study of the safety and activity of formulated IL-12 plasmid administered intraperitoneally in combination with standard neoadjuvant chemotherapy in patients with newly diagnosed advanced stage ovarian cancer.
Authors: Thaker Premal H. , Bradley William Hampton , Leath Charles A. , Gunderson Camille Catherine , Borys Nicholas , Anwer Khursheed , Musso Lauren , Alvarez Ronald David
Published: November 2019

155

Background: This study evaluated weekly intraperitoneal GEN-1, an IL-12 plasmid formulated with polyethyleneglycol-polyethyleneimine cholesterol lipopolymer, in combination with dose-dense weekly taxane (T) and carboplatinum (C) every 3 weeks in newly diagnosed epithelial ovarian, fallopian tube, or primary peritoneal cancer (EOC) patients undergoing neoadjuvant therapy (NAC). The primary objective of this study was to evaluate the tolerability and safety of GEN-1 with NAC. Secondary objectives were objective clinical response and pathological complete response at interval debulking surgery (IDB). Methods: This protocol followed a standard 3+3 design. Patients received GEN-1 at escalating dose levels from 36 mg/m2, 47 mg/m2, 61 mg/m2, to 79 mg/m2 weekly for 8 treatments with concurrent T/C. After IDB, patients did not receive further GEN-1 but continued on T/C. Patients were assessed for dose limiting toxicities (DLT) and had tumor samples at a time of diagnostic laparoscopy and IDB. Translational endpoints are yet to be analyzed. Patients are being followed for every 3 months for 2 years. Results: Fifteen patients were enrolled and 9 patients received all 8 treatments with no DLTs. The 79 mg/m2GEN-1 cohort of patients are actively being treated. Two patients were screen failures; 1 patient discontinued prior to GEN-1 administration due to port site infection. Most common related toxicities were Gr 1 nausea, fatigue, abdominal pain, and vomiting. One patient experienced Gr 2 fevers associated with GEN-1 but responded to acetaminophen and fluids. One patient did not undergo IDB due to pulmonary embolism and severe deconditioning due to underlying cancer. Of the other 8 patients undergoing IDB: 3/8 had stable disease, 4/8 had partial responses (PR), and 1/8 had complete response (CR). There was 1 pathological CR, 3 micro PR, and 3 macro PR. All patients had a ≥ 89% drop in CA-125 following GEN-1. Conclusions: Adding GEN-1 to T/C is safe and appears to be active in NAC patients with newly diagnosed EOC. Updated results to be presented at meeting. Clinical trial information: NCT02480374.
Identification of T-cell-inflamed gastric adenocarcinoma in The Cancer Genome Atlas (TCGA).
Authors: Maron Steven Brad , Luke Jason John , Hovey Raymond , Bao Riyue , Gajewski Thomas , Ji Yuan , Catenacci Daniel V.T.
Published: November 2019

16

Background: Gastroesophageal adenocarcinoma (GEC) is a significant global health problem. KEYNOTE-012 demonstrated a 22% objective response rate in patients with PD-L1 expressing GEC that received pembrolizumab. A subset of patients (pts) tumors express a T cell “inflamed” (TCI) phenotype, which can be measured using an IFN-γ-based immune signature and may prove more predictive of clinical benefit. Using a 160 gene RNA-Seq immune expression profile, we sought to characterize the molecular environments of TCI versus non-TCI GEC patients in The Cancer Genome Atlas (TCGA). Methods: 395 GEC pts with primary tumors in TCGA were clustered into TCI, non-TCI, and intermediate subtypes using both unsupervised hierarchical and K-means clustering (k = 3). Molecular characteristics were categorized using data acquired via CbioPortal and the UCSC Xena repository. Only non-silent somatic mutations and copy number variations (CNVs) reaching GISTIC2 -2 or +2 thresholds were considered. Statistical comparisons were performed using chi-square, ANOVA, and t-test. Results: The TCI subtype contained patients from all TCGA-defined subtypes - EBV-associated (56%), MSI-high (16%), chromosomal unstable (6%), and genomically stable (27%). No significant differences were seen between TCI and non-TCI for tumor site or stage. Mutations in PTEN, PIK3CA, CDH1, and RHOA were more frequent in TCI patients. ERBB2, CCNE1, and KRAS CNVs were infrequent in TCI patients as were PDE4D deletions ( p< 0.05 ). TCI tumors had higher expression of both co-inhibitory (PD-1, PD-L1/L2, CD28/80, BTLA, LAG3) and co-stimulatory (CD137/40/27, OX40, GITR, ICOS) checkpoint molecules ( p< 10-7). Total mutation burden was no different between TCI and non-TCI pts when excluding MSI-high pts nor when assessing MSI-high alone. Conclusions: The IFN immune phenotype encompassed GEC patients from all TCGA subsets. Correlation of clinical outcome with checkpoint blockade is necessary to confirm these molecular associations and the independent predictive utility of this immune-profile stratification.
Immunotherapy and survival in stage IV melanoma: A National Cancer Database (NCDB) analysis.
Authors: Ahmed Aabra , Krishnan Mridula , Siu Margaret , Walters Ryan W , Silberstein Peter T.
Published: November 2019

160

Background: Interleukin-2 (IL-2) has been the predominant immunotherapy used for stage IV melanoma for the past decades, with newer immunotherapy drugs approved within the past five years. By using the National Cancer Database (NCDB), our study aims to be the largest to determine the impact of immunotherapy on patients with stage IV melanoma. To examine only IL-2 and interferon-alpha, only patients diagnosed prior to 2011 were used. Methods: 10,771 patients diagnosed with stage IV melanoma between 2004-2010 were identified with sufficient data. 1,009 of these patients were given immunotherapy. Chi-square analysis was used to determine demographic differences between those with and without immunotherapy. Between-therapy survival differences were estimated by the Kaplan-Meier method and associated log-rank tests; Tukey-Kramer adjusted p < .05 indicated statistical significance. Results: Patients with immunotherapy had a median survival of 17.4 months, while those without immunotherapy had a median survival of 7.1 months. 27% of patients given immunotherapy were alive three years post diagnosis, compared to only 14% of patients who were not given immunotherapy. Immunotherapy patients had a greater percentage of those with private insurance (65.3% vs. 37.7%, p<0.001), an income greater than $63,000 (40.5% vs. 31.9%, p<0.001), and no comorbidities (87.3% vs. 77.5%, p<0.001) compared to patients not given immunotherapy. Conclusions: Patients with immunotherapy had nearly a 2.5-fold increase in median survival compared to patients not provided immunotherapy. Compared to those without immunotherapy, immunotherapy patients lived in more wealthy and educated zip codes, and had more private insurance. [Table: see text]
Sipuleucel-T (sip-T) induced lytic CD8+ T cell responses to target antigens in men with hormone-sensitive and castration-resistant prostate cancer (CRPC).
Authors: Antonarakis Emmanuel S. , Quinn David I. , Kibel Adam S. , Petrylak Daniel Peter , Vu Tuyen , Campogan Dwayne , Chang Nancy N. , Trager James B. , Sheikh Nadeem Anwar , Drake Charles G.
Published: November 2019

162

Background: Sip-T is an FDA-approved immunotherapy for patients (pts) with asymptomatic or minimally symptomatic metastatic CRPC. Sip-T is manufactured from autologous peripheral blood mononuclear cells cultured with the immunogen PA2024, a fusion antigen of prostatic acid phosphatase (PAP) conjugated to granulocyte macrophage colony-stimulating factor. After sip-T, antibody and T cell responses to PA2024 and/or PAP correlate with improved survival. To further elucidate the mechanism of sip-T–induced immune responses, we evaluated the proliferative and lytic ability of PA2024- and PAP-specific CD8+ T cells. Methods: Mononuclear blood cells were labeled with the membrane dye carboxyfluorescein succinimidyl ester (CFSE) and cultured with PA2024 or PAP. In vitro proliferative and lytic CD8+ (cytotoxic T lymphocyte [CTL]) T cell responses to these antigens were evaluated by flow cytometry. For proliferation, progressive dilution of CFSE was measured. For CTL activity, the loss of intracellular granzyme B (GzB), indicating exocytosis of this apoptosis-mediating enzyme, was assessed. Samples were from 2 sip-T clinical trials STAND (NCT01431391) and STRIDE (NCT01981122), hormone-sensitive and CRPC pts, respectively. Results: Six wk after sip-T administration, CD8+ PAP- and PA2024-specific responses were observed (n=14 pts assessed). The magnitude of PA2024-specific CD8+ proliferative responses was greater than that for PAP-specific responses. CD8+ T cells from a subset of pts who exhibited PA2024- and/or PAP-specific proliferative responses were assessed for lytic ability. After in vitro antigen stimulation, CTL activity in all evaluated samples (n=14, PA2024; n=13, PAP) was demonstrated by a significant decrease (p<0.05) in intracellular GzB relative to a no-antigen control. Conclusions: Sip-T induced CD8+ CTL proliferation against the target antigens PAP and PA2024. Moreover, antigen-specific CTL activity provides the first direct evidence that sip-T can induce tumor cell lysis. These antigen-specific CD8+ lytic abilities were observed within 6 wk following sip-T, suggesting rapidly generated immune responses. Clinical trial information: NCT01431391; NCT01981122.
Cytokine profile of sipuleucel-T (sip-T) in differentiating reactivation of latent immunity from de novo immune responses.
Authors: Drake Charles G. , Petrylak Daniel Peter , Antonarakis Emmanuel S. , Kibel Adam S. , Chang Nancy N. , Vu Tuyen , Campogan Dwayne , Haynes Heather , Trager James B. , Sheikh Nadeem Anwar , Quinn David I.
Published: November 2019

163

Background: Sip-T is an FDA-approved immunotherapy for treating patients (pts) with asymptomatic or minimally symptomatic metastatic castration-resistant prostate cancer (mCRPC). It is manufactured from autologous peripheral blood mononuclear cells (PBMCs) cultured with PA2024, a fusion of prostatic acid phosphatase (PAP) and granulocyte macrophage colony-stimulating factor. Survival of sip-T–treated mCRPC pts correlates with immune responses to PA2024 and/or PAP. PA2024- or PAP-specific CD4+ and CD8+ T cell proliferation and cytokine production and release were assessed to better understand sip-T–induced T cell responses. Methods: Pts with biochemical recurrence or mCRPC were from sip-T trials (NCT01431391, NCT01981122). PBMCs collected at baseline through 6 mo post–sip-T were cultured in vitro and stimulated with PA2024 or PAP. CD4+ and CD8+ T cells were assessed (n=19) for proliferation and intracellular IL-2 and IFN-γ. The cytokine profile was confirmed in supernatant with a meso scale discovery assay. P<0.10 was statistically significant. Results: Compared with baseline, PA2024-specific proliferating CD4+ and CD8+ T cells had increased intracellular IL-2 and IFN-γ levels at wk 6 and mo 6, with a similar trend for PAP-specific proliferating T cells (Table 1). Compared with unstimulated controls, a significant >2-fold increase in PA2024-stimulated IL-2 and IFN-γ in supernatant was observed at wk 6 and mo 6 over baseline (p<0.001). PAP-stimulated IL-2 and IFN-γ supernatant levels increased over baseline and were significantly elevated for IFN-γ at wk 6 (p<0.10). Conclusions: Sip-T therapy generated a de novo PA2024-specific T cell response, as indicated by the cytokine release profile. The PAP-stimulated cytokine profile suggests that pre-existing immunity with terminally differentiated T cells are expanded. Thus, sip-T reactivated an anti-PAP response in memory T cells, thereby overcoming immunosuppressive mechanisms in PC. Clinical trial information: NCT01431391; NCT01981122. [Table: see text]
Feasibility study of multiple HLA-class IA restricted peptide vaccines (KRM-19) for metastatic triple negative breast cancer (TNBC).
Authors: Toh Uhi , Saku Shuko , Iwakuma Nobutaka , Okabe Mina , Shichijo Shigeki , Yamada Akira , Itoh Kyogo , Akagi Yoshito
Published: November 2019

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Background: Our previous phase II clinical trial has indicated that the therapeutic selected personalized peptide vaccines (PPV) were effective for boosting anticancer immunity and the immune response after PPV was associated with the clinical outcome as a prognostic factor for metastatic breast cancer (mBC). Based on the data from the PPV studies, we have conducted an early phase II study to evaluate the safety and the efficacy of a new regimen using multiple peptide vaccines (KRM-19) for pts with metastatic TNBC. Methods: KRM-19 consisted of 19 mixed peptides which were chosen from the previously reported 31 PPVs according to their anti-tumor immunologic effect and safety profile for mBC pts. All patients had histologically confirmed measurable ER-PgR-Her2- mBC and their human leukocyte antigen (HLA)-A molecules should be each of -A2, A3, A11, A24, A26, A31, or A33. KRM-19 (19mg/ml) was administrated subcutaneously in order with schedule of every week for a total of 6 doses. The concurrent conventional chemo- and/or endocrine therapy was not permitted for the combination, but was available for post-study treatment. Measurement of peptide-specific cytotoxic T lymphocyte (CTL) and IgG responses were conducted before and after vaccination. Levels of IgG reactive to each of the 19 peptides in the pre- and post-vaccination plasma were measured using the LUMINEX system at every 6 vaccinations. Peptide-specific CTL responses were examined by INF-ɤ ELISPOT. Clinical response was evaluated by RECIST criteria. Results: 10 patients with refractory metastatic TNBC received all 6 vaccines and 10/10 patients experienced Grade 1-2 skin reaction at injection site, 5/10 pts had Grade 2-3 liver function disorder, 3/10 and 1/10 pts had Grade 2 bone marrow suppression and nausea, respectively. The clinical responses were assessed in 8 pts included 1 PR case, 4 SD cases, and 3 PD cases. The evaluation of other clinical and immunologic data is underway. Conclusions: Subcutaneous KRM-19 vaccine administration was safe and resulted in a 12.5% objective response and 62.5% clinical benefit rate in TNBC pts, warranting further larger scale study. Clinical trial information: UMIN000014616.
Effect of RELT on regulation of the threshold of T-cell activation during sterile inflammation.
Authors: Kim Seon-Hee , Choi Beom K. , Han Chungyong , Kwon Byoung S.
Published: November 2019

25

Background: Receptor expressed in lymphoid tissue (RELT) is a type I transmembrane protein, designated TNFRSF19-like, and primarily expressed in lymphoid tissues and immune cells. However, its immunological function has yet to be characterized. Methods: We developed RELT-deficient mice to examine the immunological role of RELT. The RELT–/– mice were exposed to viral and bacterial infection, chemical-induced liver injury, and tumor induction. Results: RELT–/– mice were viable and fertile, and developed normal lymphoid and myeloid cells. RELT transcripts were decreased in T cells and dendritic cells following their activation, and T-cell proliferation was enhanced in the absence of RELT in vitro. Nevertheless, we could not find significant changes in RELT–/– mice infected with virus or bacteria. However, liver injury and inflammation were significantly increased in RELT–/– mice in comparison to RELT+/+ mice after the injection of acetaminophen. Tumor growth rate was also slower in RELT–/– than RELT+/+ mice. Transfer of naïve or activated pmel-1 CD8+ T cells suppressed the growth of B16–F10 tumors and increased host CD8+ tumor-infiltrating cells more efficiently in RELT–/– than in RELT+/+ mice. In particular, naïve pmel-1 CD8+ T cells were present in increased numbers and activated forms in draining lymph nodes of RELT–/– than in RELT+/+ mice, whereas activated pmel-1 CD8+ T cells were not. Conclusions: As RELT–/– mice only showed significant differences in comparison to RELT+/+ mice in pathogen-associated-molecular-pattern-free conditions, these results provide evidence that RELT functions as a negative modulator of T cell responses in a sterile inflammation.
HER2 gene amplification and protein overexpression in uterine clear cell carcinoma and its implications in targeted immunotherapy.
Authors: Zhou Jain , Sukov William R. , Carter Jodi M , Schoolmeester J. Kenneth
Published: November 2019

29

Background: Uterine clear cell carcinoma (UCCC) is a high-grade endometrial carcinoma. The current treatment is hysterectomy with post-operative chemotherapy and/or radiation. The 5-year disease free survival remains dismal for UCCC with 65% for early-stage and 54% for advanced stage disease. In addition, UCCC may be more resistant to chemotherapy or radiation therapy than the endometrioid subtype. The aim of the current study is to investigate the HER2 gene amplification status in UCCC and its role for targeted therapy in UCCC. Methods: Twenty-nine cases of UCCC were retrieved from surgical pathology archives of Mayo Clinic at Rochester between 2011 and 2015. All cases except one case were hysterectomy specimens. The blocks contain the most characteristic morphology of UCCC were selected and corresponding paraffin sections were subjected to fluorescent in situ hybridization for amplification of HER2 gene (Hercept, Abbott Molecular) and parallel immunohistochemical (IHC) study. Results: A total of 9 (of 29; 31%) UCCCs showed HER2 amplification and 4 (of 29; 14%) were considered equivocal for HER2 amplification by FISH. A total of 3 (10%) tumors showed 3+ HER2 overexpression while 11 (38%) UCCCs showed 2+ HER2 overexpression, 9 (31%) showed 1+ expression with the remaining cases showing no expression of HER2. Importantly, we observed significant intratumoral heterogeneity with regard to HER2 expression. Comparing the results of IHC with HER2 gene status as determined by FISH, 2 (66%) of the 3 cases that showed 3+ HER2 expression also showed amplification for HER2 by FISH, while 1 (33%) was equivocal for HER2 amplification. Of the 11 tumors that showed 2+ HER2 expression, 6 (55%) were amplified by FISH and 1 (9%) was equivocal. Conclusions: This is the largest number of UCCC cases that has been studied on the HER2 amplification and corresponding protein overexpression. Our results indicate that the HER2 overexpression is common in UCCCs and is frequently associated with HER2 amplification. These results also suggest that targeted adjuvant therapy with trastuzumab-based immunotherapy should be evaluated in patients with UCCC showing HER2 protein overexpression or HER2 gene amplification.
Immune characterization of the programmed death receptor pathway in high-risk prostate cancer.
Authors: Alanee Shaheen Riadh , Baas Wesley , Gershburg Svetlana , Dynda Danuta , Delfino Kristin , Robinson Kathy , Yearley Jennifer
Published: November 2019

40

Background: Programmed Death 1 (PD-1) is a T cell inhibitory receptor critical to a major immunomodulation pathway which has been implicated in tumor evasion of immune response. PD-1 and its ligand PD-L1 have been found to be expressed in many tumor types, and this expression has led to the development of drugs targeting the PD-1 pathway. The goal of this study was to understand the expression of PD-1, and PD-L1 in high grade prostate cancer tissue, and correlate the expression with disease and patients characteristics. Methods: Immunohistochemistry for PD-1 (CD279), PD-L1 (B7-H1), and CD3 was performed on prostatectomy/biopsy tissue samples taken from 25 men with high grade (Gleason 8-10) prostate cancer using anti-PD-L1 clone 22C3 (Merck) and anti-PD-1 clone NAT105 (Cell Marque) . CD3 was used as a specific marker for T cell infiltration. Charts were then retrospectively reviewed for patient and disease characteristics including age at diagnosis, race, Gleason score, prostatic specific antigen (PSA) level at diagnosis, number of positive cores at biopsy, volume of tissue on biopsy and/or prostatectomy involved by cancer, clinical TNM stage, pathologic TNM stage, biochemical recurrence, or metastasis. Statistical analyses were done to correlate these patient and disease characteristics with PD-1, PD-L1, and CD3 expression. Results: A score of 3-5 on the semi-quantitative 0-5 score was deemed “high” expression whereas a score of 0-2 was deemed “low” expression. Of the 25 samples, 2 (8%) scored high for PD-1 expression, 2 (8%) scored high for PD-L1 expression, and 18 (72%) scored high for CD3 expression. With independent t-tests there was found to be no statistically significant correlation between any of the variables we collected and expression of PD-1, PD-L1, or CD3. Conclusions: We found an overall low expression of PD-1 and PD-L1, and a concurrent high expression of CD3+ T cells in high risk prostate cancer tissue. No significant correlations were made between expression of PD-1, PD-L1, or CD3 and patient and disease characteristics. The elevated T cell content in high risk prostate cancer is particularly interesting with the continued emergence of new drugs focused on enhancing efficacy of the immune response in cancer.