bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract A64: Chronic allergic contact dermatitis, A potent tumor promoter of Marjolin's ulcer
Authors: Demehri Shadmehr , Cunningham Trevor J. , Hurst Eva A. , Schaffer Andras , Sheinbein David M. , Yokoyama Wayne M.
Published: October 2015

Allergic contact dermatitis (ACD) is a well-recognized adverse event associated with implantable medical devices that contain allergenic materials like nickel. However, other cutaneous consequences of such chronic exposures are not well understood. Herein, we report a clinical case of Marjolin's ulcer, an invasive squamous cell carcinoma (SCC) developing at the site of chronic inflammation from a previously unreported cause: chronic ACD caused by an orthopedic implant. To determine whether chronic ACD promotes skin carcinogenesis, we utilized a standard murine model of contact hypersensitivity. The chronic application of 1-Fluoro-2,4-dinitrobenzene (DNFB) to carcinogen-treated skin led to the development of papillomas and aggressive SCC. The DNFB-driven chronic ACD was marked by type 2 inflammation that mediated the skin carcinogenesis. Importantly, we found similar tumor-promoting inflammation surrounding the SCC in our patient. Our findings demonstrate that chronic ACD caused by constant exposure to an allergen can promote tumorigenesis at skin sites with pre-existing cancer-initiated cells. This has great implication for the monitoring of implantable devices that are placed in the proximity of the skin and highlights the importance of patch testing prior to the placement of such devices.

Citation Format: Shadmehr Demehri, Trevor J. Cunningham, Eva A. Hurst, Andras Schaffer, David M. Sheinbein, Wayne M. Yokoyama. Chronic allergic contact dermatitis, A potent tumor promoter of Marjolin's ulcer. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A64.
Abstract B07: Characterization of T-cell therapy enhancing oncolytic adenoviruses with human cytokine expression
Authors: Havunen Riikka , Parviainen Suvi , Siurala Mikko , Hemminki Akseli
Published: October 2015

Adenovirus vectors are the most commonly used gene therapy vectors in clinical trials. Administrated systemically or intratumorally the virus is capable of inducing immune responses against the tumor per se, but this effect can also be amplified by arming the viruses with immune system stimulating factors. Moreover, we have discovered that adenoviruses enhance adoptive T-cell therapy in a potent manner. Our previous data also shows that the most promising factors to stimulate immune system are interleukin (IL) -2 and Tumor Necrosis Factor alpha (TNFα). IL-2 is a commonly used cytokine in treating malignant melanoma and renal cell carcinoma. However, systemic IL-2 can in some cases lead to severe side effects. Like IL-2, TNFα activates immune cells, but it is also known for its anti-tumor properties and it is for example capable of promoting tumor cell death by apoptosis and necrosis. Arming oncolytic viruses with cytokines results in long lasting, high level expression locally but low level systemically. Currently, we are developing oncolytic adenoviruses with expression of one or both of the above mentioned human cytokines. The viruses have an Ad5/3 chimera that is generated through replacement of the Ad5 fiber knob with the Ad3 knob while the Ad5 shaft and tail are retained. The chimera has proven to have an improved cancer cell transduction as well as antitumor efficacy. In addition, the viral genome has an E2F promoter and a 24-basepair deletion at the E1A domain, which makes it selective to retinoblastoma protein defective cells (most tumor cells). The viral particles are produced in A549 lung adenocarcinoma cell line. The genomes are sequenced and the functionality of the viruses and the cytokines proven with various methods. The secretion of cytokines is tested with ELISA and by using indicator cell lines. The functionality of the viruses is studied by infecting different cells lines with variable amount of viral particles and measuring the proportion of surviving cells with MTS-assay. Our data proves oncolytic and cytokine production capability of Ad5/3-E2F-D24-hTNFα/hIL2 adenoviruses. The results indicate that these viruses can be used in further investigations in vivo.

Citation Format: Riikka Havunen, Suvi Parviainen, Mikko Siurala, Akseli Hemminki. Characterization of T-cell therapy enhancing oncolytic adenoviruses with human cytokine expression. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B07.
Abstract B13: Prostate cancer immunotherapy development using tumor-specific CAR design
Authors: Prima Victor , Kusmartsev Sergei , Vieweg Johannes
Published: October 2015

Prostate cancer that has progressed to metastatic disease remains largely untreatable. One of the most promising modalities for treatment of advanced cancer is immunotherapy based on adoptive transfer of T lymphocytes. It was demonstrated that T lymphocytes expressing tumor-specific chimeric artificial receptor (CAR) exert potent anti-tumor activity upon adoptive transfer into tumor-bearing host. Recent studies identified Prostate-specific membrane antigen (PSMA) and ligands for NK cell –activating receptors (e.g. NKG2D ligands) as the targets primarily expressed on tumor cells but absent on most normal tissues. We generated two sets of chimeric DNA constructs: CAR-PSMA and CAR-NKG2D, that can be used for transduction of human T cells. Both of them consist of an antigen-recognizing domain bound to the transmembrane and intracellular domains of various signal-transducing and/or cell-activating molecules and a signaling molecule. Extracellular antigen-recognizing domain of CAR-PSMA consists of single chain anti-PSMA antibody (scFv) while in another construct set it is human NKG2D receptor. Intracellular CD3 ξ-chain functions as a signaling molecule triggering T-cell activation. Co-stimulatory proteins CD28, CD40L or OX40L which can compensate for the lack of co-stimulation from the target tumor cell are either fused to CD3 ξ-chain as intracellular domains or are expressed on cell surface from bi-cistronic constructs.

Upon transduction of human T cells with these constructs, transduced cells presented surface expression of the chimeric scFv anti-PSMA or NKG2D, respectively. In vitro experiments demonstrated specific binding activity and cytotoxicity of human T lymphocytes expressing anti-PSMA or NKG2D CARs against prostate cancer cells which creates the basis for a viable prostate cancer immunotherapy development.

*VP and SK contributed equally to the project.

Citation Format: Victor Prima, Sergei Kusmartsev, Johannes Vieweg. Prostate cancer immunotherapy development using tumor-specific CAR design. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B13.
Abstract B22: A chimeric antigen receptor (CARs) based upon a killer immunoglobulin-like receptor (KIR) triggers robust cytotoxic activity in solid tumors
Authors: Wang Enxiu , Wang Liang-Chuan , Tsai Ching-Yi , Bhoj Vijay , Albelda Steven , Moon Edmund , Milone Michael C.
Published: October 2015

Chimeric antigen receptors (CARs) based upon a single chimeric molecule bearing an antigen binding domain linked in cis to the cytoplasmic domains of CD3ζ and costimulatory receptors CD28 or 4-1BB provide a potent method for engineering T cell cytotoxicity towards tumors. We describe a simple chimeric multichain receptor based upon a killer immunoglobulin-like receptors (KIRs) normally expressed by natural killer (NK) cells and T cells. Constructed by fusing a single chain variable fragment (scFv) to the transmembrane and cytoplasmic domain of a KIR, we show that a KIR-based CAR targeting mesothelin (SS1-KIR) triggers antigen-specific cytotoxic activity and cytokine production that is comparable to CD3ζ-based CARs with antigen-induced proliferation that is independent of additional costimulation. Using a xenograft model of mesothelioma resistant to T cell immunotherapy, we further demonstrate that a KIR-based CAR targeting mesothelin exhibits more potent anti-tumor activity compared with T cells bearing mesothelin-specific CD3ζ-based CARs with costimulation despite in vivo persistence of the latter CAR-modified T cells. Evaluation of tumor infiltrating lymphocytes demonstrate that KIR-based CAR+ T cells show resistance to acquired hypofunction within the tumor microenivornment compared with CD3ζ-based CARs with costimulatory receptor domains. The ability of a KIR-based CAR to induce regression of a tumor in which second generation CD3ζ-based CARs show limited activity supports the future clinical evaluation of a KIR-based CAR in mesothelioma and other tumors.

Citation Format: Enxiu Wang, Liang-Chuan Wang, Ching-Yi Tsai, Vijay Bhoj, Steven Albelda, Edmund Moon, Michael C. Milone. A chimeric antigen receptor (CARs) based upon a killer immunoglobulin-like receptor (KIR) triggers robust cytotoxic activity in solid tumors. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B22.
Abstract B33: Endoplasmic reticulum stress enhances the antigen-specific T cell immune responses and therapeutic antitumor effects generated by therapeutic HPV vaccines
Authors: Lee Sung Yong , Shim JaeJeong , Ho Kang Kyung , Lee Kye Young , Hung Chien-Fu , Wu T.-C.
Published: October 2015

Endoplasmic reticulum stress has a profound effect on cancer cell proliferation and survival, and also has the capacity to activate cells of the adaptive immune system. Multimodality treatments that combine conventional cancer therapies with antigen-specific immunotherapy have emerged as promising approaches for the control of cancer. However, it is not well known whether ER stress inducing agents can influence the efficacy of tumor antigen targeted vaccines. We used previously developed therapeutic human papillomavirus (HPV) DNA vaccines encoding calreticulin (CRT) linked to HPV-16 E7 antigen (CRT/E7) vaccine and ER stress inducing agent, 3-bromopyruvate in a preclinical model for its potential to enhance E7-specific CD8+ T cell immune responses as well as antitumor effects against E7-expressing tumor (TC-1 cells). In vitro assay, luciferase-expressing TC-1 tumor cells were treated with 3-bromopyruvate. E7-specific CD8+ T cells were added to evaluate the cytotoxicity. The degree of cytotoxic T lymphocytes-mediated killing of the tumor cells was measured with the IVIS Luminescence Imaging System. We also determined the levels of ER stress markers in 3-bromopyruvate treated TC-1 cells. In vivo assay, TC-1 tumor-bearing mice were treated with 3-bromopyruvate (10mg/kg, intraperitoneal injection) and/or CRT/E7 DNA vaccine (30μg/mouse). The tumor mass size and survival were evaluated. Treatment of TC-1 cells with 3-bromopyruvate induced significant in vitro cytotoxicity. We found that treatment with 3-bromopyruvate led to upregulation of ER stress markers (CHOP, GRP78) in E7-expressing TC-1 tumor cells. More importantly, treatment with combination of 3-bromopyruvate and CRT/E7 DNA vaccine led to improved antigen-specific CD8+ T cell immune responses as well as therapeutic antitumor effects in TC-1 tumor-bearing mice. Thus, our data indicate that 3-bromopyruvate can enhance therapeutic HPV vaccine potency in generating improved antigen-specific immune responses and antitumor effects. These findings have important implications for future clinical translation.

Citation Format: Sung Yong Lee, JaeJeong Shim, Kang Kyung Ho, Kye Young Lee, Chien-Fu Hung, T.-C. Wu. Endoplasmic reticulum stress enhances the antigen-specific T cell immune responses and therapeutic antitumor effects generated by therapeutic HPV vaccines. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B33.
Abstract B37: VSV-GP: A vaccine vector and oncolytic virus
Authors: Muik Alexander , Kimpel Janine , Tober Reinhard , Urbiola Carles , Laer Dorothee von
Published: October 2015

Oncolytic viruses have recently shown significant clinical benefits in several cancers. Interestingly, in addition to debulking the tumor by direct oncolysis, they were found to act as a cancer vaccine by setting free tumor antigen in the presence of virus-induced danger signals, which have an adjuvant effect. However, as oncolytic viruses induce neutralizing antibodies that inactivate the virus, oncolytic viruses are not delivered efficiently to cancer tissue upon repeated systemic application. Thus, repeated applications of oncolytic viruses have been restricted to intratumoral injections, which clinically has not been convincingly effective in metastatic disease.

We generated the oncolytic virus VSV-GP (Muik et al., Cancer Res, 2014) that is highly potent, lacks toxicity even at high doses (so far tested in mice and rabbits), can be produced at high titers and, above all, does not induce neutralizing antibodies readily. Thus, VSV-GP is unique among the oncolytic viruses in that it can be applied repeatedly without loosing efficacy. VSV-GP is highly effective leading to cure in several cancer mouse models, i.e. malignant glioma, melanoma, prostate cancer and ovarian cancer. In addition to oncolysis, VSV-GP induced a sound anti-tumor immune response and acted synergistically with other cancer immunotherapeutics. Moreover, VSV-GP was shown to be a highly promising vaccine vector and is the only replication competent viral vaccine vector that efficiently boosts the immune response upon repeated application. VSV-GP is thus a highly versatile replication competent viral vector platform for vaccination and cancer therapy.

Citation Format: Alexander Muik, Janine Kimpel, Reinhard Tober, Carles Urbiola, Dorothee von Laer. VSV-GP: A vaccine vector and oncolytic virus. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B37.
Abstract B63: The Macrophage Migration Inhibitory Factor promotes breast cancer metastasis through interaction with the host immune response
Authors: Balogh Kristen , Cross Janet
Published: October 2015

The Macrophage Migration Inhibitory Factor (MIF) is an inflammatory cytokine that has been well-characterized as a mediator of numerous inflammatory disease processes. We have previously demonstrated that expression of MIF from the primary tumor is required for spontaneous pulmonary metastasis in the 4T1 murine model of breast cancer. MIF is overexpressed in many cancer types, including breast cancer, and the degree of MIF expression correlates with tumor aggressiveness. We therefore sought to understand the link between primary tumor expression of MIF and increased pulmonary metastasis. As a cytokine, we hypothesized that MIF would function by controlling some aspect of the interaction between the tumor and the host immune system. This was supported by our observation that MIF expression was no longer required for metastasis in an immunodeficient SCID/bg host. Using flow cytometry, we discovered that MIF expression in the primary tumor increased the abundance of a subset of myeloid-derived suppressor cells (MDSCs) in the tumor microenvironment. Furthermore, systemic depletion of MDSCs from tumor-bearing mice prevented spontaneous metastasis, suggesting that the MIF-induced increase in MDSCs is likely mediating the enhanced metastasis. Our subsequent observations suggest that MIF and MDSCs may be cooperating not only in the primary tumor, but also at sites of metastasis. Based on these results, we hypothesize that MIF and MDSCs are functioning to promote formation of a “pre-metastatic niche” in the lung. Ongoing studies aim to determine if MIF expression in the primary tumor is actively remodeling lung tissue to promote recruitment and/or differentiation of myeloid cells in the lungs. Metastatic disease is the most common cause of cancer mortality in patients with breast cancer, and gaining a better understanding of how MIF and MDSCs are working to induce metastasis could lead to significant therapeutic advancement for these patients.

Citation Format: Kristen Balogh, Janet Cross. The Macrophage Migration Inhibitory Factor promotes breast cancer metastasis through interaction with the host immune response. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B63.
Abstract B65: Dissecting the tumor myeloid compartment reveals rare activating antigen presenting cells, critical for T cell immunity
Authors: Broz Miranda , Binnewies Mikhail , Boldajipour Bijan , Nelson Amanda , Pollock Joshua , Erle David , Barczak Andrea , Rosenblum Michael , Daud Adil , Barber Diane , Amigorena Sebastian , Veer Laura J. van 't , Sperling Anne , Wolf Denise , Krummel Matthew F.
Published: October 2015

Incoming tumor-reactive cytotoxic T lymphocytes (CTL) are modulated by innate immune cells of the tumor microenvironment and we have used intravital live-imaging to characterize the predominant T cell-APC interactions there. To further hone in on the identities of the APC populations, we have extensively examined myeloid subset diversity across multiple spontaneous (GEMM) and ectopic models. Importantly, as a functional counterpart to the abundant tumor infiltrating macrophages, we have identified a small population of rare tumoral Dendritic Cells capable of robust antigen processing and T cell stimulation. We have identified the specific lineage requirements of these cells, dissecting the essential cytokines driving each tumor APC population. In addition we have found that T-cell dependent immune clearance relies upon these rare tumoral Dendritic Cells. Thus, through the dissection of the tumor myeloid compartment we have revealed a rare, yet highly stimulatory and targetable DC subset that could be exploited therapeutically to enhance anti-tumor immune responses locally.

This work is supported by the Genentech Foundation and the Margaret A. Cunningham Immune Mechanisms in Cancer Research Fellowship Award.

Citation Format: Miranda Broz, Mikhail Binnewies, Bijan Boldajipour, Amanda Nelson, Joshua Pollock, David Erle, Andrea Barczak, Michael Rosenblum, Adil Daud, Diane Barber, Sebastian Amigorena, Laura J. van 't Veer, Anne Sperling, Denise Wolf, Matthew F. Krummel. Dissecting the tumor myeloid compartment reveals rare activating antigen presenting cells, critical for T cell immunity. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B65.
Abstract B67: A big role in tumor immune microenvironment of a small non-coding RNA: microRNA-200
Authors: Chen Limo , Goswami Sangeeta , Yi Xiaohui , Byers Lauren , Diao Lixia , Roybal Jonathon , Ungewiss Christin , Peng David , Wang Jing , Wistuba Ignacio , Chen Lieping , Ullrich Stephen , Heymach John , Kurie Jonathan , Qin Xiao-Feng , Gibbons Don
Published: October 2015

Immunosuppression of tumor-infiltrating lymphocytes (TIL) is a common feature of advanced epithelial tumors, but its biological basis has remained obscure. In addition, anti-PD-1/PD-L1 treatment has produced encouraging clinical responses, but it is still unclear which patients are likely to benefit. Thus, successful therapeutic translation will require a thorough understanding of mechanisms of treatment resistance and predictive biomarkers to select patients.

Using multiple lung cancer mouse models (spontaneous and transplant models), we demonstrate a molecular link between epithelial-to-mesenchymal transition (EMT) and intra-tumoral CD8+ T cell immunosuppression, two key drivers of malignant tumor progression. We show that microRNA-200 (miR-200), a cell-autonomous suppressor of EMT and metastasis, targets PD-L1, a checkpoint inhibitor of CD8+ T cell immunity. Moreover, ZEB1, an EMT activator and transcriptional repressor of miR-200, relieves miR-200 repression of PD-L1 on tumor cells, leading to CD8+TIL dysfunction and metastasis. These findings were further supported by robust correlations between the EMT score, miR-200 levels and PD-L1 expression in multiple human lung cancer datasets: The Cancer Genome Atlas (TCGA) lung adenocarcinoma dataset and the Profiling of Resistance patterns and Oncogenic Signaling Pathways in Evaluation of Cancers of the Thorax (PROSPECT) project at MD Anderson Cancer Center.

In addition to revealing a link between EMT and T cell dysfunction, these findings also show that ZEB1 promotes metastasis through a heretofore unappreciated cell non-autonomous mechanism, and they suggest that subgroups of patients in whom malignant progression is driven by EMT activators may respond to treatment with PD-L1 antagonists. Our data also suggest that EMT represents an important biomarker to select the patients who may benefit from immune checkpoint blockade agents and other immunotherapies in lung cancer and possibly a broader range of other cancers.

Citation Format: Limo Chen, Sangeeta Goswami, Xiaohui Yi, Lauren Byers, Lixia Diao, Jonathon Roybal, Christin Ungewiss, David Peng, Jing Wang, Ignacio Wistuba, Lieping Chen, Stephen Ullrich, John Heymach, Jonathan Kurie, Xiao-Feng Qin, Don Gibbons. A big role in tumor immune microenvironment of a small non-coding RNA: microRNA-200. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B67.
Abstract B68: BAMLET in combination with the TLR7 agonist R848 protects against melanoma in a murine model
Authors: Dyck Lydia , Harte Níal , Mok Kenneth Hun , Mills Kingston H.G.
Published: October 2015

BAMLET, a bovine α-lactalbumin - oleic acid complex, is a novel and promising candidate for the treatment of cancer because of its capacity to specifically kill tumor cells while leaving normal cells and tissues relatively unharmed. While recent studies focused on direct tumoricidal effects of BAMLET, we investigated the potential of BAMLET to modulate anti-tumor immune responses, in combination with the TLR7 agonist R848, which stimulates innate and adaptive immunity.

Consistent with previous studies, BAMLET induced killing of B16 melanoma cells in vitro. Interestingly, we demonstrated for the first time that BAMLET also has immunostimulatory effects. BAMLET enhanced R848-induced dendritic cell (DC) maturation and IL-12, TNF and IL-6 production, which are crucial for the induction of adaptive immune responses against tumors. Moreover, DCs stimulated with BAMLET and R848 were capable of activating CD4 T cells in vitro. Treatment of tumor bearing mice with either BAMLET or R848 alone conferred some protection against tumor growth. However, treatment with a combination of BAMLET and R848 conferred a high level of protection, with 57% of mice completely rejecting the tumor. We found a higher frequency of DCs and IFNγ-secreting T cells in the tumor draining lymph nodes and spleens of mice treated with the combination of BAMLET and R848, compared with untreated mice or mice treated with either BAMLET or R848 alone.

Our data are consistent with a role for BAMLET in directly killing tumor cells, which release antigens that are taken up by DCs, but suggest that BAMLET can also promote maturation and secretion of T cell polarizing cytokines by DCs, especially when co-activated through a TLR. These activated DCs promote induction of IFNγ-secreting CD4 T cells that have anti-tumor activity. In conclusion, we demonstrated that BAMLET and R848 are a promising combination therapy against cancer as they target tumor cells directly, but also activate anti-tumor immune responses.

Citation Format: Lydia Dyck, Níal Harte, Kenneth Hun Mok, Kingston H.G. Mills. BAMLET in combination with the TLR7 agonist R848 protects against melanoma in a murine model. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B68.
Abstract B75: CCL2 promotes epithelial-mesenchymal transition (EMT) in prostate cancer cells via downregulation of dual-specificity phosphatase 22
Authors: Lin Wen-Jye , Izumi Kouji , Lin Showping , Ho Hui-Min
Published: October 2015

CCL2 is a member of the CC chemokine superfamily that plays a crucial role in mediating macrophage recruitment and prostate cancer (PCa) metastasis. We have discovered that inhibiting the function of androgen receptor (AR) in prostate cancer cells results in the upregulation CCL2 and promotes disease progression by recruiting macrophages, activating signal transducer and activator of transcription 3 (STAT3), and by favoring the epithelial-to-mesenchymal transition. However, the molecular switches controlling CCL2 downstream signaling in PCa cells are not well understood. Here, we established an in vitro cell migration screening system using transwell migration chambers, and identified a higher amount of CCL2 in migrating PCa cells, establishing a link between CCL2 and a subpopulation of PCa cells. Importantly, we found that CCL2 expression correlated with the EMT marker, N-cadherin in migrating PCa cells. Importantly, CCL2 stimulation in PCa cells enhanced the expression levels of several EMT markers and induced STAT3 activation. In addition, CCL2 stimulation in PCa cells induced the downregulation of dual-specificity phosphatase 22 (DUSP22), which has been shown to act as a negative regulator of the IL-6-induced STAT3 activation through a direct interaction with STAT3. This supports that CCL2-induced DUSP22 downregulation in PCa cells as a potential mechanism to promote EMT. Therefore, we believe CCL2 may promote PCa metastasis in an autocrine manner through promotion of EMT in PCa cells with more aggressive motility behaviors. Targeting of the CCL2-STAT3 axis may offer new approaches to suppress the multifaceted support (immunosuppression, macrophage recruitment, and angiogenesis) of CCL2 for PCa cells in complex tumor microenvironment.

Citation Format: Wen-Jye Lin, Kouji Izumi, Showping Lin, Hui-Min Ho. CCL2 promotes epithelial-mesenchymal transition (EMT) in prostate cancer cells via downregulation of dual-specificity phosphatase 22. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B75.
Abstract B84: Live imaging reveals hypoxia to be a limiting factor in CTL-mediated tumor rejection
Authors: Shakhar Guy , Feferman Tali , Manaster Yoav , Hutzler Anat
Published: October 2015

Background: One of the obstacles to successful immunotherapy by CTLs is thought to be poor vascularization of tumors that leads to sub-optimal tumor infiltration. In contrast, the effects of disrupted vascularization and the hypoxia it introduces on CTL migration and function within tumors have not been investigated.

Methods: To study whether tumor vascularity can shape anti-tumor CTL responses, we combined immunohistochemistry of human melanoma samples with dynamic two-photon imaging within B16-OVA tumors in live mice.

Results: In patients, CD8 T cells were twofold denser around peripheral vessels than around deep vessels, and fourfold denser than in avascular areas. This pattern was clearest in poorly-infiltrated tumors. Correspondingly, in mice, OT-I CTLs preferentially infiltrated B16-OVA tumors through peripheral ICAM1+ vessels. CTLs remained initially at their entry areas, crawling vigorously only within 50 µm of flowing blood vessels. Gradually, CTLs formed dense perivascular swarms, producing locally-high CTL-to-target ratios at their advancing front. These swarms advanced through the tumors, leading to patchy elimination of tumor cells. Target killing was antigen-specific, contact-dependent and perforin-mediated, and required several CTLs to engage one target cells to be effective. Several findings suggest that CTL confinement to perivascular areas is oxygen-dependent: occlusion of intratumoral blood vessels resulted in immediate arrest of CTLs. Immunohistology indicated that CTLs avoided hypoxic tumor areas. Hypoxia and mitochondrial uncoupling interfered with CTL migration in vitro and, finally, highly vascular tumors grown in a matrix containing pro-angiogenic factors were more easily rejected after CTL injection, displayed delayed growth in untreated mice but were not affected in immunodeficient mice.

Conclusions: We suggest that CTLs depend on oxygen for intratumoral locomotion and cytolysis. Incomplete vascularization promotes patchy infiltration and tumor escape; tumor immunotherapy may thus benefit from normal and extensive tumor angiogenesis

Citation Format: Guy Shakhar, Tali Feferman, Yoav Manaster, Anat Hutzler. Live imaging reveals hypoxia to be a limiting factor in CTL-mediated tumor rejection. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B84.
Abstract B86: The NF-κB p65 and p50 homodimer cooperate with pSTAT1 to synergistically activate iNOS transcription
Authors: Simon Priscilla , Sharman Sarah , Lu Chunwan , Yang Dafeng , Paschall Amy , Liu Kebin
Published: October 2015

Inducible nitric oxide synthase (iNOS) metabolizes L-arginine to produce NO that was originally identified in myeloid cells as a host defense mechanism against pathogens. Recent studies, however, have revealed that iNOS is often induced in both tumor and myeloid cells in the tumor microenvironment. More intriguingly, compelling experimental data have shown that iNOS promotes tumor development in certain cellular context and suppress tumor development under other cellular conditions. The molecular mechanisms underlying this contrasting function of iNOS is unknown. Because iNOS is often induced by inflammatory signals, it is therefore likely that the opposite functions of iNOS might be controlled by the inflammatory signaling pathways. We show here that proinflammatory IFN-γ and NF-κB signals synergistically induce iNOS expression in human colon cancer cells. We further demonstrated that NF-κB directly binds to the NOS2 promoter to regulate iNOS expression. Although the IFN-γ signaling pathway or the NF-κB signaling pathway alone is insufficient to induce iNOS expression in myeloid cells, IFN-γ and NF-κB synergistically induce iNOS expression in myeloid cells. Furthermore, we determined that IFN-γ up-regulates IRF8 expression to augment NF-κB induction of iNOS expression. More interestingly, we observed that the p65/p65 and p50/p50 homodimers, not the canonical p65/p50 heterodimer, directly binds to the nos2 promoter to regulate iNOS expression in myeloid cells. Our results thus determine that the IFN-γ-induced IRF8 acts in concert with the NF-κB p65/p65 and p50/p50 homodimers to regulate iNOS expression.

Citation Format: Priscilla Simon, Sarah Sharman, Chunwan Lu, Dafeng Yang, Amy Paschall, Kebin Liu. The NF-κB p65 and p50 homodimer cooperate with pSTAT1 to synergistically activate iNOS transcription. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B86.
Abstract B90: Poly (lactide)-based nanoparticles by free-radical dispersion polymerization: Fabrication, characterization and in vitro studies
Authors: Adesina Simeon K. , Akala Emmanuel O.
Published: October 2015

We have used the macromonomer method to prepare crosslinked, paclitaxel-loaded PLA-PEG (stealth) nanoparticles using free-radical dispersion polymerization. The nanoparticles were optimized using statistical D-optimal mixture design. Nanoparticle fabrication with poly(lactide) is mainly carried out by the dispersion of preformed polymers which makes it difficult to attach targeting moieties to the surface of the nanoparticle and to produce crosslinked networks; hence we designed a new approach based on dispersion polymerization. Confirmation of nanoparticle synthesis was by scanning electron microscopy. Particle size and size distribution and the zeta potential of the optimized formulation were determined using Zetasizer nano Zs. The release profile of paclitaxel-loaded nanoparticles was determined by high performance liquid chromatography and encapsulated drug was released over 7 days. In vitro cytotoxicity studies were carried out using the CellTiter®glo luminescent cell viability assay in MCF7, MDA-MB-231 (breast cancer) and SK-OV-3 (ovarian cancer) cell lines. The cytotoxicity assay shows that the blank nanoparticle is biocompatible with no toxicity for the duration of the assay compared to medium-only treated controls. Further, the paclitaxel-loaded nanoparticle formulation exhibits similar cytotoxicity compared to free drug in solution against the cancer cell lines tested. In vitro intracellular localization of nanoparticle by confocal microscopy also demonstrated that the nanoparticles are rapidly internalized by MCF-7 cancer cells within one hour probably by nonspecific endocytosis. The stealth nanoparticles are suitable for the design of controlled delivery systems for bioactive agents.

Citation Format: Simeon K. Adesina, Emmanuel O. Akala. Poly (lactide)-based nanoparticles by free-radical dispersion polymerization: Fabrication, characterization and in vitro studies. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B90.
Abstract IA01: Molecular basis of T cell exhaustion: Insights for immunotherapy
Authors: Wherry E. John
Published: October 2015

T cell exhaustion is common during chronic infections, cancer and prevents optimal immunity. Exhausted T cells are defined by the loss of ability to perform effector functions efficiently, low proliferative capacity and poor survival following antigen stimulation. In addition, it has become clear that exhausted T cells co-express multiple inhibitory receptors that negatively regulate their function. Indeed, receptors such as PD-1 have become major targets of clinical immunotherapies in cancer and infectious disease aimed at re-invigorating exhausted T cells. Our work has recently defined transcriptional networks of T cell exhaustion and has focused on the role of key transcription factors, including T-bet and Eomesodermin (Eomes) in controlling the sustainability and terminal differentiation of exhausted T cell populations. We have found a key role for T-bet in sustaining a progenitor pool of exhausted CD8 T cells during chronic infection, while the related transcription factor Eomes governs terminal differentiation. Our new data indicates that PD-1, the major current target of immunotherapy approaches dramatically regulates these subsets of exhausted T cells and modulating PD-1 signals may have long-term impact on lineage dynamics of exhausted T cell populations. One key observation from these studies is that a key surrogate of reivingoration of exhausted CD8 T cell responses is a population of EomesHi PD-1Hi exhausted CD8 T cells. The use of this subset of CD8 T cells as a biomarker is explored. Ultimately, a more precise molecular understanding of T cell exhaustion should lead to novel and more robust clinical interventions to reverse exhaustion in settings of persisting infections and cancer.

Citation Format: E. John Wherry. Molecular basis of T cell exhaustion: Insights for immunotherapy. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr IA01.
Abstract IA19: Curative potential of T-cell immunotherapy for cancer
Authors: Rosenberg Steven A.
Published: October 2015

Adoptive cell transfer (ACT) immunotherapy for patients with metastatic melanoma using autologous tumor infiltrating lymphocytes (TIL) mediated a 56% objective response rate including 20% of patients with durable complete regression ongoing from 6.7 to 10.3 years. Administration of autologous TIL to 9 patients with HPV induced metastatic cervical cancer mediated objective responses in 3 patients including 2 complete regressions that are ongoing beyond a year.

The ideal targets for ACT are the unique mutations that occur in cancers. Using deep exomic sequencing a technique has been developed to identify any cancer mutation, presented on any of the patient's MHC molecules, that gives rise to reactive T cells. We recently reported the successful application of this approach to treat a patient with a metastatic bile duct cancer. Since virtually all cancers contain mutations this approach is now being vigorously studied to expand the current reach of cancer immunotherapy to common epithelial cancers.

Genes encoding conventional alpha-beta T cell receptors (TCR) or chimeric antigen receptors (CAR) can be efficiently transduced into autologous lymphocytes though choosing suitable targets expressed on the cancer is critical to avoid toxicities to essential normal tissues. ACT using CARs to target the CD19 molecule present on normal B cells as well as on the great majority of B cell lymphomas and leukemias was first reported to successfully treat a patient with refractory lymphoma in 2010 in a patient who remains progression-free at five years. Durable complete and partial responses have been seen in patients with chemotherapy refractory indolent and aggressive large B cell lymphomas. Of 9 heavily pretreated patients with large cell lymphomas receiving ACT, 4 patients have had complete regressions, 3 ongoing from 9 to 22 months and 2 additional patients have had partial responses.

Cancer-testes antigens such as NY-ESO-1 and MAGE-A3 are expressed during fetal development and in 10 to 80% of cancers from multiple tissues but are often not expressed in adult normal tissues. ACT targeting NY-ESO-1 resulted in a 67% response rate in 15 treated patients with refractory synovial cell sarcoma and a 53% response rate in 19 patients with melanoma including durable complete regressions. Shared mutations that are unique to an individual cancer type also represent excellent targets for cell transfer immunotherapy and we are conducting a trial using a CAR targeting the EGFRvIII mutation expressed in about 40% of patients with high grade glioblastoma

Citation Format: Steven A. Rosenberg. Curative potential of T-cell immunotherapy for cancer. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr IA19.
Neural Correlates of Schizophrenia Negative Symptoms: Distinct Subtypes Impact Dissociable Brain Circuits
Authors: Shaffer Joseph J. , Peterson Michael J. , McMahon Mary Agnes , Bizzell Joshua , Calhoun Vince , van Erp Theo G.M. , Ford Judith M. , Lauriello John , Lim Kelvin O. , Manoach Dara S. , McEwen Sarah C. , Mathalon Daniel H. , O''Leary Daniel , Potkin Steven G. , Preda Adrian , Turner Jessica , Voyvodic Jim , Wible Cynthia G. , Belger Aysenil
Published: October 2015

Background: The negative symptoms of schizophrenia include deficits in emotional expression and motivation. These deficits are stable over the course of illness and respond poorly to current medications. Previous studies have focused on negative symptoms as a single category; however, individual symptoms might be related to separate neurological disturbances. We analyzed data from the Functional Biomedical Informatics Research Network dataset to explore the relationship between individual negative symptoms and functional brain activity during an auditory oddball task. Methods: Functional magnetic resonance imaging was conducted on 89 schizophrenia patients and 106 healthy controls during a two-tone auditory oddball task. Blood oxygenation level-dependent (BOLD) signal during the target tone was correlated with severity of five negative symptom domains from the Scale for the Assessment of Negative Symptoms. Results: The severity of alogia, avolition/apathy and anhedonia/asociality was negatively correlated with BOLD activity in distinct sets of brain regions associated with processing of the target tone, including basal ganglia, thalamus, insular cortex, prefrontal cortex, posterior cingulate and parietal cortex. Conclusions: Individual symptoms were related to different patterns of functional activation during the oddball task, suggesting that individual symptoms might arise from distinct neural mechanisms. This work has potential to inform interventions that target these symptom-related neural disruptions.
Recent Trend of Hepatitis E Virus Infection in Chiba Area, Japan: 3 of 5 Cases with Rheumatoid Arthritis
Authors: Kanda Tatsuo , Yasui Shin , Nakamura Masato , Arai Makoto , Sasaki Reina , Haga Yuki , Wu Shuang , Nakamoto Shingo , Okamoto Hiroaki , Yokosuka Osamu
Published: October 2015

Hepatitis E virus (HEV) infection is an emerging health concern in developing and developed countries, such as Japan. Five cases have recently been diagnosed as hepatitis E. Of interest, 3 of them had rheumatoid arthritis (RA), although a previous study demonstrated a lack of association between HEV and RA. One of the other patients developed autoimmune hepatitis and was successfully treated with corticosteroids approximately 150 days after the diagnosis of hepatitis E. In RA patients with liver dysfunction, the presence of HEV infection should be evaluated immediately because these patients are often relatively old. Further investigation of the association between HEV and autoimmune hepatitis is needed.
Lugares públicos como potencialidades: uma leitura urbana do centro histórico de São Paulo
Authors: Rupf Lilian Dazzi Braga , Queiroga Eugênio Fernandes
Published: October 2015

Este trabalho propõe um ensaio de leitura urbana a fim de apreender relações entre o espaço urbano produzido e as vivências cotidianas no Centro Histórico de São Paulo, através da observação empírica do presente como espessura que remete ao passado e futuro. Ao contrário de alguns discursos, não há uma redução de sua vida pública, o que há é uma mudança nesta, que adquire maior complexidade: novos usos e dinâmicas ocupam seus lugares públicos. Hoje a área central tem como herança um rico sistema de espaços livres que protagoniza grandes momentos da vida pública paulistana; estes, independentemente do perfil e da apropriação dos usuários, são fortemente utilizados.
Políticas para gerenciamento da mídia exterior na paisagem urbana: um paralelo entre os casos de São Paulo e Florianópolis
Authors: Casarin Vanessa , Santiago Alina Gonçalves
Published: October 2015

A mídia exterior está presente de forma ostensiva nas grandes cidades contemporâneas e tem sido utilizada de forma desordenada na paisagem, causando prejuízos para as cidades e seus usuários e contribuindo para a “poluição visual” urbana. Este artigo tem por objetivo traçar um paralelo entre duas experiências distintas no enfrentamento dessa situação, a da cidade de Florianópolis, que implantou um projeto intitulado Renovar, em 1993, no perímetro central de caráter histórico, e a recente iniciativa implementada na cidade de São Paulo, através da Lei Cidade Limpa, que abrangeu a totalidade do município. A discussão se dá com base em dados coletados através de entrevistas realizadas com os profissionais envolvidos na implantação dos referidos projetos. Os resultados das experiências abordadas através da reflexão sobre a prática foram confrontados e apontaram para uma dificuldade no gerenciamento da mídia exterior visível do espaço público, apesar de ela ser apenas um dos aspectos que compõem o cenário visual urbano, e que, no entanto, tem forte impacto na imagem da cidade. Nesse sentido, também ficou evidente a necessidade de o município controlar e gerenciar a mídia exterior em conjunto com os demais aspectos que contribuem com a qualidade visual urbana.
DLL4/Notch1 and BMP9 Interdependent Signaling Induces Human Endothelial Cell Quiescence via P27
KIP1
and Thrombospondin-1
Authors: Rostama Bahman , Turner Jacqueline E. , Seavey Guy T. , Norton Christine R. , Gridley Thomas , Vary Calvin P.H. , Liaw Lucy
Published: October 2015

Objective—

Bone morphogenetic protein-9 (BMP9)/activin-like kinase-1 and delta-like 4 (DLL4)/Notch promote endothelial quiescence, and we aim to understand mechanistic interactions between the 2 pathways. We identify new targets that contribute to endothelial quiescence and test whether loss of
Dll4

+/
−

in adult vasculature alters BMP signaling.

Approach and Results—

Human endothelial cells respond synergistically to BMP9 and DLL4 stimulation, showing complete quiescence and induction of
HEY1
and
HEY2
. Canonical BMP9 signaling via activin-like kinase-1-Smad1/5/9 was disrupted by inhibition of Notch signaling, even in the absence of exogenous DLL4. Similarly, DLL4 activity was suppressed when the basal activin-like kinase-1-Smad1/5/9 pathway was inhibited, showing that these pathways are interdependent. BMP9/DLL4 required induction of P27
KIP1
for quiescence, although multiple factors are involved. To understand these mechanisms, we used proteomics data to identify upregulation of thrombospondin-1, which contributes to the quiescence phenotype. To test whether Dll4 regulates BMP/Smad pathways and endothelial cell phenotype in vivo, we characterized the vasculature of
Dll4

+/
−

mice, analyzing endothelial cells in the lung, heart, and aorta. Together with changes in endothelial structure and vascular morphogenesis, we found that loss of Dll4 was associated with a significant upregulation of pSmad1/5/9 signaling in lung endothelial cells. Because steady-state endothelial cell proliferation rates were not different in the
Dll4

+/
−

mice, we propose that the upregulation of pSmad1/5/9 signaling compensates to maintain endothelial cell quiescence in these mice.

Conclusions—

DLL4/Notch and BMP9/activin-like kinase-1 signaling rely on each other’s pathways for full activity. This represents an important mechanism of cross talk that enhances endothelial quiescence and sensitively coordinates cellular responsiveness to soluble and cell-tethered ligands.