bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract A81: Adoptively transferred Chondromodulin 1/HLA-A:0201 allo-restricted T cell receptor transgenic CD8+ T cells are well tolerated and cause partial regression in a patient with disseminated Ewing Sarcoma
Authors: Thiel Uwe , Kirschner Andreas , Einspieler Ingo , Burdach Stefan
Published: March 2017

Background: Patients with Ewing sarcoma (ES) metastatic to bone and bone marrow (BM) do not survive irrespective of therapy. We assessed toxicity, viability, BM homing and anti-tumor activity of adoptively transferred HLA-A*0201/chondromodulin 1 (CHM1) allo-restricted T cell receptor (TCR) transgenic CD8+ T cells in an ES patient with refractory disease including marrow.

Patient and Methods: An twelve years old HLA-A2+ boy was diagnosed with ES of the right femur. After treatment according to the Euro-Ewing 2008 protocol he suffered early relapse 11 months after diagnosis. For relapse treatment he received MetaEICESS 2007 including consolidation with haploidentical stem cell transplantation (allo-SCT) from his HLA-A2- mother. Before and shortly after transplant the patient relapsed twice. Thereafter he suffered from progressive disease under combined rescue radiochemotherapy. Within one week after the last progression, he received 1 x 106/kg (1st transfer, d0) and 8.2 x 106/kg CHM1319-T cell receptor (TCR) transgenic donor CD8+ T cells (2nd transfer, d32; in combination with ipilimumab and nivolumab), respectively. A patient derived ES cell line was isolated.

Results: CHM1319-TCR transgenic donor T cells showed specific in vitro lysis of the patient's ES cell line. One week after 1st transfer, CHM1319-TCR transgenic T cells stained CHM1319 multimer positive in BM but negative in peripheral blood (PB). After 2nd transfer, BM of posterior iliac spines as well as PB stained CHM1319 multimer positive at a high rate and RT-PCR and flow-cytometric CD99+/CD45- ES phenotype as well as partial PET-signal reduction were noted. Five further metastatic bone/ marrow compartments showed significant PET signal regression. The patient never developed GvHD symptoms and died of disease 10 weeks after 1st transfer due to resistant lung metastasis.

Conclusions: CHM1319-TCR transgenic T cells home to affected BM and cause partial disease regression. These cells may proliferate in vivo for at least four weeks without causing GvHD.

Citation Format: Uwe Thiel, Andreas Kirschner, Ingo Einspieler, Stefan Burdach. Adoptively transferred Chondromodulin 1/HLA-A:0201 allo-restricted T cell receptor transgenic CD8+ T cells are well tolerated and cause partial regression in a patient with disseminated Ewing Sarcoma. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr A81.
Abstract B02: Immune events in the sentinel lymph node in melanoma predict local and systemic response to disease
Authors: Essner Richard , Gong Ke-Wei , Kaufman David Z. , Gangi Alexandra , Sim Myung Shin
Published: March 2017

Background: The sentinel node (SN) has become the standard diagnostic tool for staging cutaneous melanoma. With the recent development of anti-CTLA-4 and anti-PD-1 antibodies as immunotherapies we hypothesized that the early immune events in melanoma maybe first observed in the SN and be related to the response to disease.

Methods: 81 SN from 79 patients were evaluated with microarray technology, qPCR, routine H&E & IHC. We identified 10 immune genes that are present by qPCR in SNs from a random section of the SN. Statistical analysis assessed the utility of these markers to predict tumor-positive SN and survival.

Results: Of the 79 patients, 48 (61%) were men. Median age was 59 (range 6-95 years). Primary melanomas were most commonly from the trunk (51%) and extremities (40%). The median thickness of the primaries was 0.95mm. 13% of the patients had ulcerated primaries. 29% of the primary tumors had at least 1 mitosis/mm2 . 9 (11%) patients had SN metastasis, as determined by conventional H&E and IHC. The incidence of SN positivity was directly related to tumor thickness; the higher the T-stage the greater percentage of SN positivity: 2% T1, 19% T2, 20% T3 and 60% T4 melanoma. 33% of ulcerated primaries and 30% with increased mitotic rate had a tumor-positive SN. The microarray did not identify specifc immune related genes when comparing tumor-positive to tumor-negative SN. We then evaluated the gene expression of CD80, CD86, CD28, CD40, IDO, Arginase, IL-2, IL-10, PD-1 and CTLA-4 by qPCR. By univariate analyses: patient age (p<0.001), primary site (<0.001), decreasing gene expression of CTLA-4 (p<0.001) and PD-1 (p<0.001) in SN were predictive of SN positivity. Multivariate analyses confirmed the down-regulation of CTLA-4 and PD-1 gene expression in the SN were predictive of metastases in the SN and the AUC values of predicting SN positivity were 0.808 and 0.731 respectively . The positive and negative predictive values of the tests were 100 and 89.5%. Further the down-regulation of CTLA-4 and PD-1 gene expression in the SN correlated with diminished survival, while the up-regulation of IL-10 correlated with poor survival.

Conclusions: Our results demonstrate the significance of CTLA-4, PD-1 and IL-10 gene expression in the SN of melanoma patients. The expression of these genes may be early events in the immune dysfunction that occurs with melanoma metastases.

Citation Format: Richard Essner, Ke-Wei Gong, David Z. Kaufman, Alexandra Gangi, Myung Shin Sim. Immune events in the sentinel lymph node in melanoma predict local and systemic response to disease. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr B02.
Abstract B06: Targeting M2-Tumor Associated Macrophages (M2-TAMs) in prostate cancer
Authors: Zarif Jelani C. , Hernandez James R. , Yang Weiming , Zhang Hui , Pienta Kenneth J.
Published: March 2017

Prostate cancer is a leading cause of cancer-related deaths of men in the U.S., and in the past year, over 30,000 men died from this disease. While localized prostate cancer is highly treatable by surgical resection and radiation, cancer that has metastasized remains incurable. Alternatively activated macrophages, also known as M2-macrophages, primarily scavenge debris and in the process, promote angiogenesis and wound repair. M2-macrophages are phenotypically similar to M2 tumor-associated macrophages (M2-TAMs) have been reported to associate with solid tumors such as prostate cancer to facilitate epithelial to mesenchymal transition (EMT), tumor invasiveness, metastasis, and resistance to therapy. As an invasive species within the tumor microenvironment, this makes M2-TAMs an ideal therapeutic target in prostate cancer. The purpose of this project is to develop novel therapeutics that will directly target M2-TAMs for destruction and subsequently attenuate prostate tumor growth, progression, and metastasis. Our hypothesis is to determine if targeting of M2-TAMs by using enriched surface antigens that are targeted by antibody-drug-conjugates (ADCs), be an effective therapy for lethal prostate cancer while simultaneously eliciting an immune response. To identify novel surface antigens expressed on M2-macrophages, we developed a novel method of creating homogenous populations of human macrophages from CD14+ monocytes in vitro. Our homogenous M1 macrophages secrete pro-inflammatory cytokines and our M2 macrophages secrete anti-inflammatory cytokines as well as VEGF. We then performed solid-phase extraction of N-linked glycopeptides (SPEG) followed by liquid chromatography-tandem Mass Spectrometry (LC-MS/MS) on our homogenous macrophage populations. We discovered six novel peptides that are enriched exclusively on M2-macrophages relative to M1 macrophages and CD14+ monocytes. Lastly, we determined if these surface antigens, found enriched on M2 macrophages, were also expressed in human metastatic castrate-resistant prostate cancer (mCRPC). Using mCRPC tissues from rapid autopsies supplied by the Departments of Urology and Surgical Pathology, we were able to determine M2-macrophage infiltration by using immunohistochemistry and flow cytometry. The studies described here outline a method of altering the tumor immune microenvironment. By identifying specific markers on M2-TAMs, we predict that this method of targeting will provide a better prognosis for patients who have been diagnosed with lethal prostate cancer.

Citation Format: Jelani C. Zarif, James R. Hernandez, Weiming Yang, Hui Zhang, Kenneth J. Pienta. Targeting M2-Tumor Associated Macrophages (M2-TAMs) in prostate cancer. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr B06.
Abstract B17: Validation of targeted gene expression profiling panel for identifying biomarker signatures of immunotherapy responders
Authors: Pankov Aleksandr , Sun Yongming , Ku Yuan-Chieh , Tom Warren , Zheng Jianping , Looney Timothy , Au-Yong Janice , Hyland Fiona , Mongan Ann
Published: March 2017

Cancer immunotherapy has led to an unprecedented, long-lasting response in populations susceptible to the therapies. Despite the therapeutic potential, identifying biomarkers and stratifying populations that are likely to respond has been a challenge. Gene expression profiling has previously been successfully used to stratify individuals based on survival and treatment characteristics, but there exist limitations with the prevalent technologies. In particular, full transcriptome gene expression estimates use limited biological material to measure the concentrations of tens of thousands uninformative genes and often lack the depth required to accurately measure expression levels of lowly-expressed genes. These genes may be critical to the identification of a signature associated with immunotherapy responders. To efficiently measure the expression of the key genes potentially informative of an immunotherapy response, we developed a high-throughput targeted gene expression solution measured by our RNA Ion Oncomine Immune Response Profiling panel* containing 395 genes. This panel provides information about the expression of genes involved in tumor checkpoint inhibition (including CTLA4, PD-1, PD-L1, OX-40, 4-1BB, TIM3, LAG3) and other targets such as CSF1R, and IDO1, as well as additional markers of T cell signaling pathway, interferon signaling, and markers of tumor infiltrating lymphocytes (TIL). We used publicly available TCGA data to demonstrate the need and develop a solution for a new normalization procedure that allows for accurate comparisons of samples within various cancer types. Furthermore, we verified a linear and unbiased estimate of fold change in our assays across mixing concentrations of a cell-line titration experiment. Finally, by achieving a high correlation (r > .99) of technical replicates, along with robust expression estimation even at low input amounts (10 ng RNA), our panel offers a valuable solution for biomarker research in cancer immunotherapy.

*For research use only. Not for use in diagnostic procedures.

Citation Format: Aleksandr Pankov, Yongming Sun, Yuan-Chieh Ku, Warren Tom, Jianping Zheng, Timothy Looney, Janice Au-Yong, Fiona Hyland, Ann Mongan. Validation of targeted gene expression profiling panel for identifying biomarker signatures of immunotherapy responders. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr B17.
Abstract B25: Visualization of immune response to Hepatitis B vaccination by in vivo mouse imaging
Authors: Kim Ha , Hong Keejong , Ahn Byung Cheol , Yum Jung Sun , Youn Hyewon
Published: March 2017

By taking advantage of bioluminescence imaging to monitor luciferase expressing splenocytes from the luciferase transgenic mouse, we visulaized the enhancement of immune response against hepatitis B virus (HBV) vaccine with adjuvants. To visualize antigen in vivo, large hepatitis B virus antigen (L-HBsAg) was labeled with radioiodine (125I). B6 mice were intramuscularly vaccinated. The localization of 125I-L-HBsAg was monitored for 5 weeks using animal SPECT/CT. To monitor the immune response, the luciferase expressing splenocytes were injected intravenously into immunized B6 mice. Bioluminescence signals from splenocytes were measured by IVIS 100 system. Localization of inoculated L-HBsAg was successfully monitored using animal SPECT/CT. L-HBsAg was lasted for 5 weeks and diminished. In addition, the injected splenocytes were successfully visualized in immunized mice, homing to spleen within 30 minutes and were accumulated in lymph nodes within 5 hours. Accumulation of splenocytes at vaccination site was observed within 24 hours. Moreover, after 10 hours, mouse vaccinated with antigen and two adjuvants showed 4 times more accumulation of splenocytes at vaccination site compared to mouse vaccinated with antigen only. Six days later, mouse vaccinated with antigen and two adjuvants showed 1.7-4.73 fold increased luciferase intensity of splenocytes at spleen, lymph nodes and vaccination site compared to mouse vaccinated with antigen only. In conclusion, in vivo real-time bioluminescent monitoring of splenocytes homing and proliferation against vaccination successfully provides efficiency of adjuvants. Our imaging system can be used for evaluation of efficacy of vaccination by enhanced the proliferation and activation of splenocytes near the vaccination site.

Note: This abstract was not presented at the conference.

Citation Format: Ha Kim, Keejong Hong, Byung Cheol Ahn, Jung Sun Yum, Hyewon Youn. Visualization of immune response to Hepatitis B vaccination by in vivo mouse imaging. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr B25.
Abstract B50: Synergistic antitumor efficacy in mice with immunotherapy regimens combining recombinant live-attenuated Listeria with immune checkpoint inhibitors
Authors: Deng Weiwen , Liu Weiqun , Hudson Thomas , Rae Chris S. , Lemmens Ed , Desbien Anthony , Hanson Bill , Lauer Pete , Dubensky Thomas W. , Leong Meredith
Published: March 2017

Development of effective and durable antitumor immunity requires the activation, expansion and maintenance of function of tumor antigen-specific effector T cells. The mechanism of action of recently FDA approved monoclonal antibody (mAb) therapies which target immune checkpoints such as PD-1 and CTLA-4 and uncouple inhibitory pathways from the activation of antigen-specific T cells underlines the critical requirement for antigen-specific priming to elicit potent and long-lasting anti-tumor immunity. However, immune checkpoint blockade results in significant clinical benefit largely in malignancies with high mutational burden, where the tumor itself can initiate T cell priming, but is less effective when used as a single agent among cancers with lower mutational burden, associated typically with a low level of lymphocyte infiltration into the tumor microenvironment (TME). We are evaluating clinical immunotherapy regimens combining immune checkpoint blockade with recombinant live-attenuated double-deleted strains of the intracellular bacterium Listeria monocytogenes (LADD), based on a hypothesis that effective immunotherapy will result from agents that in combination re-polarize the TME to facilitate immune effector cell function, prime functional tumor-specific T cells in the appropriate context, and block inhibitory signaling pathways. Here we show that tumor antigen-expressing LADD therapy enhanced CD8+ T cell effector function, resulting in significant tumor eradication in several preclinical mouse models. LADD treatment in addition induced favorable changes in the TME, as shown by enhanced CD8+ T effector function, recruitment of antigen presenting cells and reduction of regulatory T cells. Treatment regimens combining LADD-based immunotherapy with PD-1 immune checkpoint blockade significantly enhanced antitumor efficacy in CT26, 4T1 and MC38 tumor models. Together these data support the rationale for integrating LADD-based immunotherapy into clinical regimens with immune checkpoint blockade on the basis that TME modification and priming of tumor Ag-specific T cells significantly enhances the activity of mAb therapies blocking T cell inhibitory pathways.

Citation Format: Weiwen Deng, Weiqun Liu, Thomas Hudson, Chris S. Rae, Ed Lemmens, Anthony Desbien, Bill Hanson, Pete Lauer, Thomas W. Dubensky, Jr., Meredith Leong. Synergistic antitumor efficacy in mice with immunotherapy regimens combining recombinant live-attenuated Listeria with immune checkpoint inhibitors. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr B50.
Abstract B64: A high-throughput screen identifies fulvestrant as an enhancer of immune-mediated cytotoxicity of mesenchymal-like lung cancer cells
Authors: Hamilton Duane H. , Mathews-Griner Lesley , Keller Jonathan M. , Hu Xin , Southall Noel , Marugan Juan , David Justin M. , Ferrer Ferrer , Schlom Jeffrey , Palena Claudia
Published: March 2017

The phenomenon of epithelial-mesenchymal transition (EMT) is a phenotypic switch that allows epithelial tumor cells to acquire features of mesenchymal cells, including the ability to migrate and invade and, therefore, to disseminate from the primary site. In addition to promoting tumor dissemination, various reports have demonstrated that tumor EMT also associates with the acquisition of resistance to the cytotoxic effects of chemotherapy, radiation, and some targeted therapies. In the present report we have utilized a quantitative high-throughput screening of a pharmaceutical collection of more than 2000 compounds to identify FDA-approved drugs capable of augmenting the sensitivity of mensenchymal-like lung cancer cells to immune-mediated lysis. Using this approach we have identified the estrogen receptor antagonist fulvestrant as capable of rendering mesenchymal-like lung cancer cells more susceptible to the cytotoxic effects of natural killer or antigen-specific T cells and chemotherapy. Fulvestrant-treatment of human lung carcinoma cell lines, both in vivo and in vitro, was associated with a reduction of mesenchymal features. In silico analysis of publicly available gene expression datasets revealed a novel association between the expression of mesenchymal-associated features and the expression of estrogen receptor alpha in lung adenocarcinoma patients. Although the potential role of estrogen signaling in lung cancer remains controversial, this study supports a positive association between active estrogen signaling and the promotion of a mesenchymal phenotype in lung cancer. The findings from this study form the rationale for future combinations of fulvestrant and immune-mediated therapies in the treatment of advanced lung cancer.

Citation Format: Duane H. Hamilton, Lesley Mathews-Griner, Jonathan M. Keller, Xin Hu, Noel Southall, Juan Marugan, Justin M. David, Ferrer Ferrer, Jeffrey Schlom, Claudia Palena. A high-throughput screen identifies fulvestrant as an enhancer of immune-mediated cytotoxicity of mesenchymal-like lung cancer cells. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr B64.
Abstract B81: Oncolytic adenovirus YSC-02 enhanced tumor cell killing and immune response with broad spectrums, which resulted in both TGF-β and HSP27 cooperatively becoming silenced
Authors: Han Zhezhu , Je Suyeon , Choi Hye J. , Kang Dongxu , Xu Rong , Kim Hyun S. , Song Jae J.
Published: March 2017

As combining therapeutic agents with different action mechanisms may enhance efficacy, YSC-02, an oncolytic adenovirus with multi targets was loaded with five different genes, which were designed with the expectation that different action mechanisms would cooperate. In spite of concerns regarding many APIs, YSC-02 was constructed to be an adenovirus-based anti-cancer drug. By using our own established mouse model system suitable for efficient adenoviral infection and replication, immune activity as well as survival potential could be precisely estimated for anti-cancer drug efficacy. YSC-02 was designed to decrease tumor cell survival, metastasis and to increase tumor cell death, and immune activation. It is composed of five different target genes, including one fused form and two shRNAs, but each of these genes functions is closely linked to produce the maximal antitumor effect. YSC-02 is like an organic complex designed to be applied primarily to heterogeneous liver cancer and melanoma. The greatest difficulty in developing anticancer drugs is deeply related to the existence of many genetic variations even in the same tumor tissue. However, in our study, the downregulation of HSP27 greatly increased the death potential of most tumor cell types, irrespective of differential cell death induced by TGF-β1 or TGF-β2 isotypes. The simultaneous silencing of both HSP27 and TGF-β could significantly repress the tumor survival potential of various cancer cells, which leads to most cell types of death in the complex network of cells. These combined shRNAs have also contributed to the increase of TRAIL sensitivity. Genetically modified mouse cancer cells within syngenic mice could make it possible to estimate the superiority of YSC-02 over its derivatives including GMCSF-expressing oncolytic adenovirus or GMCSF-FETZ-expressing oncolytic adenovirus. At the immunological level, NK cells or T cells were shown to be most activated in the treatment of YSC-02, which may be another contributor of anti-tumor activity. Additionally, even the highest dose of YSC-02 did not produce any toxicity in the liver.

Citation Format: Zhezhu Han, Suyeon Je, Hye J. Choi, Dongxu Kang, Rong Xu, Hyun S. Kim, Jae J. Song. Oncolytic adenovirus YSC-02 enhanced tumor cell killing and immune response with broad spectrums, which resulted in both TGF-β and HSP27 cooperatively becoming silenced. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr B81.
Abstract B86: Radiation and TGFβ blockade bring back memories in metastatic breast cancer patients
Authors: Schaue Dörthe , Xie Michael W. , Ratikan Josephine A. , Micewicz Ewa D. , Hwang Lin , Faull Kym F. , Sayre James W. , Lee Percy P. , Glaspy John A. , Demaria Sandra , Formenti Silvia C. , McBride William H.
Published: March 2017

Purpose: This is a pilot study combining focal irradiation and systemic TGFβ blockade in metastatic breast cancer. The rationale for using TGFβ blockade was to limit tumor growth and further spread as well as to curb systemic immune suppression. Combining this systemic approach with hypofractionated radiation of selective tumor metastasis aimed at vaccinating each patient in vivo with her own, relevant tumor antigens by local radiation damage and allow T cells to reach their full potential while escaping TGFβ's grip.

Experimental Design: Serial blood samples from 22 patients undergoing treatment with 1mg or 10mg Fresolimumab and Radiation at the New York University School of Medicine (n=15) and at the David Geffen School of Medicine, University of California, Los Angeles (n=7) were immunophenotyped based on flow cytometric analysis of 21 surface antigens.

Results: There were significant differences between the 1mg and 10mg groups with respect to several immune parameters, especially the rise in circulating central memory CD8s at the higher dose level (relative increase at 2 weeks 10mg vs 1mg, p=0.027). Regulatory networks responded to the 10mg treatment regime with a consistent expansion in CD4 Tregs while mMDSCs declined (ratio Tregs/mMDSC rising in 10mg vs 1mg, p=0.026). An overall survival benefit was seen in the 10mg Fresolimumab arm (median OS 64.1 weeks versus 20 weeks, p=0.015 log rank test) albeit not striking. CART analysis allowed accurate survival classification based on 2-week changes in CD8/mMDSC ratios and plasma Tryptophan (ROC AUC 0.979).

Conclusion: Inhibiting TGFβ in the context of focal irradiation seems to create a favorable systemic immune landscape that drives T cell memory differentiation while limiting myeloid suppression.

Citation Format: Dörthe Schaue, Michael W. Xie, Josephine A. Ratikan, Ewa D. Micewicz, Lin Hwang, Kym F. Faull, James W. Sayre, Percy P. Lee, John A. Glaspy, Sandra Demaria, Silvia C. Formenti, William H. McBride. Radiation and TGFβ blockade bring back memories in metastatic breast cancer patients. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr B86.
Abstract IA10: The interferon receptor pathway's role in response and resistance to PD-1 blockade
Authors: Ribas Antoni
Published: March 2017

As adaptive PD-L1 expression on cancer cells is induced by interferon gamma, if a cancer had genetic alterations that lead to inability to signal from the interferon gamma receptor then this cancer would not be protecting itself from immune attack by reactively expressing PD-L1. In this scenario, it would be useless to block PD-1:PD-L1 interactions with a therapeutic antibody and it would be a genetic mechanisms of primary resistance to PD-1 blockade therapy. We identified genetic mutations in the Janus kinases (JAK) 1 and 2 controlling signaling downstream of the interferon receptor that prevent PD-L1 upregulation on melanoma cells upon interferon exposure, which are associated with primary resistance to PD-1 blockade in patients. Whole exome sequencing of metastatic melanoma biopsies from 23 patients treated with pembrolizumab revealed a JAK1 missense mutation with chromosomal amplification in the tumour with the highest mutational load in a non-responding patient. In a second cohort of 16 patients with mismatch repair deficient colon cancer treated with PD-1 blockade, a homozygous JAK1 loss-of-function mutation was similarly found in the tumour of a patient without an objective response to PD-1 blockade.

Similar alterations could explain the cases of relapse after a long period of response to PD-1 blockade therapy, termed acquired resistance. Out of the patients with metastatic melanoma who respond to pembrolizumab, approximately 75% have responses lasting years. However, delayed tumor reappearance and progression while on continuous therapy has been noted in 25% of patients. Until now, the mechanisms of immune escape in this setting were unknown. Comparing biopsies of melanoma before starting on anti-PD-1 therapy with the tumors that relapsed months to years later, we again found new JAK1/2 loss of function mutations. Among four pairs of biopsies, two had JAK1 or JAK2 new loss of function mutations with loss of heterozygosity, and the third case had a truncating mutation in the antigen presentation gene beta-2 microglobulin (B2M). JAK1 and JAK2 truncating mutations resulted in a lack of response to interferon gamma, including insensitivity to its anti-proliferative effects on cancer cells. The B2M truncating mutation led to loss of surface expression of the major histocompatibility complex (MHC) class I.

In conclusion, analysis of patient-derived biopsies is providing information about the cellular and molecular events required for response or resistance to PD-1 blockade therapy, highlighting the critical role of intact signaling through the interferon gamma receptor.

Citation Format: Antoni Ribas. The interferon receptor pathway's role in response and resistance to PD-1 blockade. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr IA10.
Abstract IA15: Engineering combination immunotherapy agents for synergistic innate and adaptive immune attack on solid tumors
Authors: Irvine Darrell J.
Published: March 2017

Solid tumors evade destruction by the immune system through many parallel pathways of immunosuppression. Combination immunotherapies that act synergistically to recruit the breadth of the immune system's armamentarium against solid tumors provide the opportunity to counter this immunosuppressive network with a similarly complex immune attack. Many existing immunotherapeutic approaches may contain key elements to overcome the barriers presented by solid tumors, but are limited by issues of toxicity (e.g., agonist immunocytokines) or potency (e.g., therapeutic vaccines). Using both established and engineered immunotherapy reagents, we recently discovered a combination immunotherapy that recruits a diverse set of innate and adaptive immune effectors, enabling robust elimination of tumor burdens that to our knowledge have not previously been curable by treatments relying on endogenous immunity. Maximal anti-tumor efficacy required four components: a tumor antigen targeting antibody, an extended-pharmacokinetics IL-2 molecule, anti-PD-1, and a lymph node targeting peptide vaccine. This combination elicited durable cures of large established tumors in a majority of animals in multiple transplanted tumor models, and induced sustained tumor regression in an autochthonous BRrafV600E/Pten-/- melanoma model. Multiple innate immune cell subsets, CD8+ T-cells, and cross-presenting DCs were critical to successful therapy. Treatment induced high levels of intratumoral inflammatory cytokines and immune cell infiltration, enhanced antibody-mediated tumor antigen uptake, and promoted antigen spreading. These results demonstrate the capacity of the endogenous immune response to destroy large, established tumors and suggest essential characteristics of an integrated immune response capable of overcoming immunosuppressive tumors.

Citation Format: Darrell J. Irvine. Engineering combination immunotherapy agents for synergistic innate and adaptive immune attack on solid tumors. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr IA15.
Abstract IA26: Genomic correlates of response and resistance to immune checkpoint blockade
Authors: Allen Eliezer M. Van
Published: March 2017

The rise of multiple immunotherapies, such as immune checkpoint blockade, has significantly improved outcomes for multiple patient populations, including melanoma, lung adenocarcinoma, and urothelial cancers, among others. In parallel, the expansion of precision cancer medicine through robust interrogation of cancer genomes at the point of care has enhanced patient stratification for multiple treatment selection strategies and cancer types. The intersection of clinical cancer genomics and immuno-oncology has created an opportunity to learn how the tumor and germline genomes, along with transcriptomic measurements of the immune microenvironment, impact clinical immune checkpoint blockade response. Here, we describe the current landscape of tumor and microenvironmental genomic correlates of immune checkpoint blockade across cancer subtypes, and examine how emerging genomic studies of immunotherapy responders and non-responders may inform patient selection strategies for clinical trial design in these diverse clinical settings. We explore how these approaches, paired with emerging computational biology methodology, may also enable discovery of new mechanisms of selective treatment response. Finally, we examine emerging genomic data on clinical resistance to immunotherapies, from both signaling pathway and neoantigen evolution perspectives, and discuss the implications for combination treatment strategies. Overall, the convergence of comprehensive clinical genomics and immuno-oncology treatment strategies may inform precision medicine for immunotherapies across cancer types.

Citation Format: Eliezer M. Van Allen. Genomic correlates of response and resistance to immune checkpoint blockade. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr IA26.
Abstract PR10: Enhancing the efficacy of T cell-based immunotherapies using miR-155 engineered tumor-specific CD8+ T cells
Authors: Ji Yun , Hocker James , Hu Jinhui , Gautam Sanjivan , Lacey Neal , Gattinoni Luca
Published: March 2017

Lymphodepleting regimens are employed prior to adoptive immunotherapy to augment the antitumor efficacy of transferred T cells by removing endogenous homeostatic cytokine sinks. These conditioning modalities, however, are often associated with severe toxicities. We found that miR-155 enabled B16 melanoma-specific CD8+ T cells to mediate profound antitumor responses in lymphoreplete hosts that were not potentiated by immune-ablation. miR-155 enhanced T cell responsiveness to limited amounts of homeostatic γc cytokines by restraining the expression of the Akt inhibitor Ship1 and multiple negative regulators of Stat5, such as Socs1 and Ptpn2, resulting in delayed cellular contraction and sustained cytokine production. To translate these findings into new clinical trials, we tested whether the overexpression of miR-155 in human T cells would enhance the antitumor efficacy of CD19-specific chimeric antigen receptor (CD19-CAR) cells against systemic acute lymphoblastic leukemia xenografts. As a safety measure, the suicide gene, inducible caspase 9, was included in the miR-155 construct. We found that miR-155 also augmented the antitumor efficacy of T cells in this xenograft tumor model. Recapitulating our findings in mouse cells, human T lymphocytes overexpressing miR-155 showed increased Stat5 signaling and enhanced survival. Our results indicate that overexpression of miR-155 in tumor-specific T cells can be employed to increase the effectiveness of adoptive immunotherapies in a cell intrinsic manner without the need of life-threatening, lymphodepleting maneuvers.

This abstract is also being presented as Poster A82.

Citation Format: Yun Ji, James Hocker, Jinhui Hu, Sanjivan Gautam, Neal Lacey, Luca Gattinoni. Enhancing the efficacy of T cell-based immunotherapies using miR-155 engineered tumor-specific CD8+ T cells. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr PR10.
Giant Fecalith Causing Near Intestinal Obstruction and Rectal Ischemia
Authors: Nigar Sofia , Sunkara Tagore , Culliford Andrea , Gaduputi Vinaya
Published: March 2017

Fecal impaction if left untreated can lead to the hardening of stools and the formation of fecalith. Fecaliths rarely cause serious complications and are usually managed conservatively. We present this rare case of a giant fecalith causing near obstruction in an institutionalized paraplegic patient at high risk for chronic constipation and fecal impaction. This case was also unusual for causing ischemic pressure necrosis in the rectum, thereby highlighting the possible serious complications of fecalith.
The Hippo Signaling Pathway Regulates Ovarian Function via the Proliferation of Ovarian Germline Stem Cells
Authors: Ye Haifeng , Li Xiaoyan , Zheng Tuochen , Hu Chuan , Pan Zezheng , Huang Jian , Li Jia , Li Wei , Zheng Yuehui
Published: March 2017

Objective: To improve the separation, identification and cultivation of ovarian germline stem cells (OGSCs), to clarify the relationship between the Hippo signaling pathway effector YAP1 and the proliferation and differentiation of OGSCs in vitro and to identify the major contribution of Hippo signaling to ovarian function. Methods: Two-step enzymatic separation processes and magnetic separation were used to isolate and identify OGSCs by determining the expression of Mvh, Oct4, Nanog, Fragilis and Stella markers. Then, YAP1, as the main effector molecule in the Hippo signaling pathway, was chosen as the target gene of the study. Lentivirus containing overexpressed YAP1 or a YAP1-targeted shRNA was transduced into OGSCs. The effects of modulating the Hippo signaling pathway on the proliferation, differentiation, reproduction and endocrine function of ovaries were observed by microinjecting the lentiviral vectors with overexpressed YAP1 or YAP1 shRNA into infertile mouse models or natural mice of reproductive age. Results: (1) The specific expression of Mvh, Oct4, Nanog, Fragilis and Stella markers was observed in isolated stem cells. Thus, the isolated cells were preliminarily identified as OGSCs. (2) The co-expression of LATS2, MST1, YAP1 and MVH was observed in isolated OGSCs. Mvh and Oct4 expression levels were significantly increased in OGSCs overexpressing YAP1 compared to GFP controls. Consistently, Mvh and Oct4 levels were significantly decreased in cells expressing YAP1-targeted shRNA. (3) After 14-75 days of YAP1 overexpression in infertile mouse models, we detected follicle regeneration in ovaries, the activation of primordial follicles and increased birth rate, accompanied by increasing levels of E2 and FSH. (4) However, we detected decreasing follicles in ovaries, lower birth rate, and decreasing E2 and FSH in serum from healthy mice of reproductive age following YAP1 shRNA expression. Conclusion: Methods for the isolation, identification and culture of OGSCs were successfully established. Further results indicate that isolated OGSCs can specifically recognize Hippo signaling molecules and that manipulation of YAP1 expression can be used to regulate the proliferation and differentiation of OGSCs, as well as ovarian function in mice. This study suggests that the Hippo signaling pathway may represent a new molecular target for the regulation of mouse ovarian functional remodeling.
A Comprehensive TALEN-Based Knockout Library for Generating Human-Induced Pluripotent Stem Cell–Based Models for Cardiovascular Diseases
Authors: Karakikes Ioannis , Termglinchan Vittavat , Cepeda Diana A. , Lee Jaecheol , Diecke Sebastian , Hendel Ayal , Itzhaki Ilanit , Ameen Mohamed , Shrestha Rajani , Wu Haodi , Ma Ning , Shao Ning-Yi , Seeger Timon , Woo Nicole , Wilson Kitchener D. , Matsa Elena , Porteus Matthew H. , Sebastiano Vittorio , Wu Joseph C.
Published: March 2017

Rationale:

Targeted genetic engineering using programmable nucleases such as transcription activator–like effector nucleases (TALENs) is a valuable tool for precise, site-specific genetic modification in the human genome.

Objective:

The emergence of novel technologies such as human induced pluripotent stem cells (iPSCs) and nuclease-mediated genome editing represent a unique opportunity for studying cardiovascular diseases in vitro.

Methods and Results:

By incorporating extensive literature and database searches, we designed a collection of TALEN constructs to knockout 88 human genes that are associated with cardiomyopathies and congenital heart diseases. The TALEN pairs were designed to induce double-strand DNA break near the starting codon of each gene that either disrupted the start codon or introduced a frameshift mutation in the early coding region, ensuring faithful gene knockout. We observed that all the constructs were active and disrupted the target locus at high frequencies. To illustrate the utility of the TALEN–mediated knockout technique, 6 individual genes (
TNNT2, LMNA/C, TBX5, MYH7, ANKRD1
, and
NKX2.5
) were knocked out with high efficiency and specificity in human iPSCs. By selectively targeting a pathogenic mutation (
TNNT2 p.R173W
) in patient-specific iPSC-derived cardiac myocytes, we demonstrated that the knockout strategy ameliorates the dilated cardiomyopathy phenotype in vitro. In addition, we modeled the Holt–Oram syndrome in iPSC-cardiac myocytes in vitro and uncovered novel pathways regulated by
TBX5
in human cardiac myocyte development.

Conclusions:

Collectively, our study illustrates the powerful combination of iPSCs and genome editing technologies for understanding the biological function of genes, and the pathological significance of genetic variants in human cardiovascular diseases. The methods, strategies, constructs, and iPSC lines developed in this study provide a validated, readily available resource for cardiovascular research.
Cardiac Outcomes After Ischemic Stroke or Transient Ischemic Attack
Authors: Young Lawrence H. , Viscoli Catherine M. , Curtis Jeptha P. , Inzucchi Silvio E. , Schwartz Gregory G. , Lovejoy Anne M. , Furie Karen L. , Gorman Mark J. , Conwit Robin , Abbott J. Dawn , Jacoby Daniel L. , Kolansky Daniel M. , Pfau Steven E. , Ling Frederick S. , Kernan Walter N.
Published: March 2017

Background:

Insulin resistance is highly prevalent among patients with atherosclerosis and is associated with an increased risk for myocardial infarction (MI) and stroke. The IRIS trial (Insulin Resistance Intervention after Stroke) demonstrated that pioglitazone decreased the composite risk for fatal or nonfatal stroke and MI in patients with insulin resistance without diabetes mellitus, after a recent ischemic stroke or transient ischemic attack. The type and severity of cardiac events in this population and the impact of pioglitazone on these events have not been described.

Methods:

We performed a secondary analysis of the effects of pioglitazone, in comparison with placebo, on acute coronary syndromes (MI and unstable angina) among IRIS participants. All potential acute coronary syndrome episodes were adjudicated in a blinded fashion by an independent clinical events committee.

Results:

The study cohort was composed of 3876 IRIS participants, mean age 63 years, 65% male, 89% white race, and 12% with a history of coronary artery disease. Over a median follow-up of 4.8 years, there were 225 acute coronary syndrome events, including 141 MIs and 84 episodes of unstable angina. The MIs included 28 (19%) with ST-segment elevation. The majority of MIs were type 1 (94, 65%), followed by type 2 (45, 32%). Serum troponin was 10× to 100× upper limit of normal in 49 (35%) and >100× upper limit of normal in 39 (28%). Pioglitazone reduced the risk of acute coronary syndrome (hazard ratio, 0.71; 95% confidence interval, 0.54–0.94;
P
=0.02). Pioglitazone also reduced the risk of type 1 MI (hazard ratio, 0.62; 95% confidence interval, 0.40–0.96; log-rank
P
=0.03), but not type 2 MI (hazard ratio, 1.05; 95% confidence interval, 0.58–1.91;
P
=0.87). Similarly, pioglitazone reduced the risk of large MIs with serum troponin >100× upper limit of normal (hazard ratio, 0.44; 95% confidence interval, 0.22–0.87;
P
=0.02), but not smaller MIs.

Conclusions:

Among patients with insulin resistance without diabetes mellitus, pioglitazone reduced the risk for acute coronary syndromes after a recent cerebrovascular event. Pioglitazone appeared to have its most prominent effect in preventing spontaneous type 1 MIs.

Clinical Trial Registration:

URL:
http://clinicaltrials.gov
. Unique identifier: NCT00091949.
Media Studies, the Spatial Turn, and the Middle East
Authors: El-Hibri Hatim
Published: March 2017

What questions do the ‘spatial turn’ in the humanities and the social sciences pose for the study of the media and culture of the Middle East? And how might attending to the spaces and spatiality of media in the Middle East help us to better understand the historical present? This article puts Middle East and Arab media and cultural studies into dialogue with an interdisciplinary literature that considers media as spatial and geographic phenomena. I examine how the question of space has arisen or might contribute to the study of media and culture in the Middle East by examining three areas of research in which that question has emerged: the place of media in domestic and public spaces and mobilities, the representation of place and space, and the geography of media industries and media infrastructure.
Molecular systematics and phylogeography of the genus Echinops (Compositae, Cardueae–Echinopsinae): focus on the Iranian centre of diversification
Authors: MONTAZEROLGHAEM SOMAYEH , SUSANNA ALFONSO , CALLEJA JUAN ANTONIO , MOZAFFARIAN VALIOLLAH , RAHIMINEJAD MOHAMMAD REZA
Published: February 2017

A molecular phylogeny of the genus Echinops (Asteraceae, Cardueae) based on plastid and nuclear markers is presented. The study focuses on the Iranian representatives of the genus, which constitute a larger part of Echinops. The Bayesian cpDNA tree resulted in a large polytomy. However, the Bayesian ITS tree displayed a well-resolved topology more compatible with the taxonomic treatment generally accepted in Echinops. An ITS network illustrates a close relationship of haplotypes. Mapping diagnostic morphological characters onto the ITS tree revealed a high level of homoplasy among the studied taxa. Except for the life cycle, all other morphological characters evolved many times among the clades and most of them appeared to be of low taxonomical value. Based on the nucleotide polymorphism and haplotypes, the taxonomic status of some taxa was evaluated. The independent status of E. sect. Phaeochaete was confirmed by molecular analyses and morphological characters. The paraphyletic nature of E. sect. Echinops was confirmed. As for the molecular dating, our results suggest that the genus evolved in the early Miocene but the main speciation events took place in the Pliocene-Pleistocene. The biogeographic results do not support any specific ancestral area for Echinops, yet the model states that its current diversity originated in the Irano-Turanian Region and the Eastern Mediterranean region. Some taxonomic changes are suggested, in the synonymy and placement of some taxa.
Description of the female of Ceromya glaucescens Tachi & Shima (Diptera: Tachinidae) and discovery of unusual sexual dimorphism in this species
Authors: TACHI TAKUJI
Published: March 2017

Sexual dimorphism is a phenomenon in which the male and female of a species differ in features of the external morphology such as size, shape, colour, or the development of appendages. In the Diptera, stalked compound eyes, leg modifications and wing patterns are well-known examples of sexual dimorphism (McAlpine 1979; Zeil 1983; Adler & Adler 1991; Meyerrochow & Reid 1994; Wilkinson & Dodson 1996; Sivinski 1997; Baker & Wilkinson 2001; Eberhard 2002; Puniamoorthy et al. 2008). Males and females of sexually dimorphic species are often described as separate species due to the dissimilarity in external characters, thus leading to problems in identification and proper association of the sexes. In contrast to characters that are usually involved in sexual dimorphism, leg chaetotaxy is considered one of the invariable character systems, irrespective of sex, in the tribe Siphonini of the Tachinidae, and is thus widely used in keys to genera and species (O’Hara 1989; Andersen 1996). Species’ identification by DNA barcoding has been used in various groups of organisms (Hebert et al. 2003; Ratnasingham & Hebert 2013). In insects, males, usually more easily identified by morphological characters (e.g., postabdominal features) than females, are often used for barcoding. The identification of females will improve as sequence data accumulate, such as data from pairs collected in copula. In this paper, I describe sexual dimorphism in the Japanese endemic species of tachinid fly Ceromya glaucescens Tachi & Shima, 2000 of the tribe Siphonini, and use molecular and morphological data for the identification of this species. Sequence data of C. silacea (Meigen, 1824) are also included for comparison.
A new Subleuconycta (Lepidoptera: Noctuidae: Acronictinae) species from Taiwan
Authors: KISS ÁDÁM , WU SHIPHER , MATOV ALEXEI YU.
Published: March 2017

The genus Subleuconycta Kozhantshikov, 1950 belongs to the noctuid subfamily Acronictinae (Kozhantshikov 1950, Kononenko 2005, 2010) and has long been known to contain only two described species (Poole 1989, Kononenko 2010). Subleuconycta palshkovi (Filipjev, 1937) occurs in the Eastern Manchurian subregion of the Palaearctic Realm, while Subleuconycta sugii Boursin, 1962 is endemic to Taiwan.