In vivo generation of 'vaccination nodes' using injectable alginate hydrogels for cancer immunotherapy

Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Materials Science and Engineering, 2009.Cataloged from PDF version of thesis.Includes bibliographical references (p. 78-90).Despite the amount of ongoing intensive research, tumor cells have continued to outwit us in the effort to combat and prevent cancer by exerting a number of mechanisms to evade and suppress anti-tumor immune responses. The present work employs strategies to generate a synthetic extranodal immunoplatform that can harbor both exogenously provided and endogenously recruited immune cells (primed against tumor cells), at the same time providing immuno-factors that support these cells and counter immunosuppressive effects from tumors. We have developed injectable self-gelling alginate formulations for this purpose, enabling sustained release of soluble immunomodulatory factors from the gels and presentation of immobilized immunostimulatory factors inside the gels. The hydrogels injected into the back flanks of mice formed a macroporous structure that allowed easy cell infiltration and migration. Modulation of the mechanical properties of self-gelling alginate was possible by varying the number of calcium-bound microspheres in the gels. During characterization of immune responses using these hydrogels, alginate gels carrying activated dendritic cells (DCs) were shown to dramatically increase the number of T cells recruited to the local injection site. When the dendritic cells were pulsed with antigen, these 'vaccination nodes' were able to initiate an antigen-specific immune response, with some of the injected DCs migrating to the regional lymph nodes and priming cognate T cells. The activated antigen-specific T cells then migrated to the injection site and infiltrated the gels, causing an effector re-trafficking phenomenon that guided both T cells and host dendritic cells to the gels.(cont.) Taking advantage of this phenomenon, the ability of the vaccination nodes to serve as a peri-tumoral local therapy against established tumors was tested using an ovalbumin-expressing B16FO subcutaneous melanoma model. When mice bearing 7-day established small tumors (-3mm2 diameter) were immunized using alginate carrying activated DCs, a mild tumor suppression effect was observed. The anti-tumor effect was augmented by supplementing IL-15 superagonist (IL-15SA) into the gels, which caused suppression of larger tumors (-20-50mm2), treated 14 days after tumor cell inoculation, and enhanced survival of the mice. In addition to showing therapeutic benefits against established tumors, the matrix-based approach allowed analysis of cells that trafficked locally near the tumor site. The ease of encapsulating factors and the injectable, non-invasive nature of the self-gelling alginate open up possibilities for use in other tissue engineering and regenerative medicine applications.by Yuki Hori.Ph.D

Modular injectable matrices based on alginate solution/microsphere mixtures that gel in situ and co-deliver immunomodulatory factors

Biocompatible polymer solutions that can crosslink in situ following injection to form stable hydrogels are of interest as depots for sustained delivery of therapeutic factors or cells, and as scaffolds for regenerative medicine. Here, injectable self-gelling alginate formulations obtained by mixing alginate microspheres (as calcium reservoirs) with soluble alginate solutions were characterized for potential use in immunotherapy. Rapid redistribution of calcium ions from microspheres into the surrounding alginate solution led to crosslinking and formation of stable hydrogels. The mechanical properties of the resulting gels correlated with the concentration of calcium-reservoir microspheres added to the solution. Soluble factors such as the cytokine interleukin-2 were readily incorporated into self-gelling alginate matrices by simply mixing them with the formulation prior to gelation. Using alginate microspheres as modular components, strategies for binding immunostimulatory CpG oligonucleotides onto the surface of microspheres were also demonstrated. When injected subcutaneously in the flanks of mice, self-gelling alginate formed soft macroporous gels supporting cellular infiltration and allowing ready access to microspheres carrying therapeutic factors embedded in the matrix. This in situ gelling formulation may thus be useful for stimulating immune cells at desired locales, such as solid tumors or infection sites, as well as for other soft tissue regeneration applications.United States. Defense Advanced Research Projects Agency (Contract W81XWH-04-C-0139)National Institutes of Health (U.S.) (EB007280-02)National Science Foundation (U.S.) (Award 0348259

Learned spatial data partitioning

Due to the significant increase in the size of spatial data, it is essential to use distributed parallel processing systems to efficiently analyze spatial data. In this paper, we first study learned spatial data partitioning, which effectively assigns groups of big spatial data to computers based on locations of data by using machine learning techniques. We formalize spatial data partitioning in the context of reinforcement learning and develop a novel deep reinforcement learning algorithm. Our learning algorithm leverages features of spatial data partitioning and prunes ineffective learning processes to find optimal partitions efficiently. Our experimental study, which uses Apache Sedona and real-world spatial data, demonstrates that our method efficiently finds partitions for accelerating distance join queries and reduces the workload run time by up to 59.4%

Transpupillary thermotherapy for atypical central serous chorioretinopathy

Ryosuke Kawamura1,2, Hidenao Ideta1, Hideyuki Hori1, Kenya Yuki2, Tsuyoshi Uno1, Tatsurou Tanabe1, Kazuo Tsubota2, Tsutomu Kawasaki11Ideta Eye Hospital, Kumamoto, Japan; 2Keio University, School of Medicine, Department of Ophthalmology, Tokyo, JapanBackground: Central serous chorioretinopathy (CSC) has been traditionally treated with laser photocoagulation. We thought that transpupillary thermotherapy (TTT) utilizing a lower temperature than that of conventional laser photocoagulation might minimize permanent retinal and choroidal damage. Studies suggest that undesirable effects on vision due to TTT are minimal even if it is applied to foveal and/or parafoveal lesions when TTT requires a larger irradiation spot. The aim of this study was to evaluate the efficacy of TTT in the management of atypical CSC.Methods: We defined atypical CSC as bullous retinal detachment with diffuse or several leakages, severe leakage with fibrin formation under serous retinal detachment, or leakage within a pigment epithelium detachment. Eight consecutive patients with atypical CSC underwent visual acuity testing, ophthalmic examination, color photography, fluorescein angiography, and optical coherence tomography to evaluate the results of transpupillary thermotherapy. Retreatment of atypical CSC was based on ophthalmic examination, optical coherence tomography, and fluorescein angiography. TTT was performed on the leaking spots shown in fluorescein angiography, with a power of 50–250 mW, spot size of 500–1200 µm, and exposure time of 13–60 seconds to minimize retinal damage.Results: In five of eight affected eyes, serous detachments completely resolved within 1 month after the initial TTT. One eye had persistent subretinal fluid and required a second TTT treatment. Two eyes showed no resolution of CSC and were treated by conventional photocoagulation. Initial best-corrected visual acuity (BCVA) ranged from 20/600 to 20/20 (mean, 20/40; median, 20/30). Final BCVA ranged from 20/200 to 20/20 (mean, 20/25; median, 20/20). BCVA improved in all cases. Only two eyes with persistent subretinal fibrin and existing retinal pigment epithelial alternations in macular area showed limited improvement of BCVA despite the absence of subretinal exudation. The presence of retinal attachment was confirmed by optical coherence tomography in six eyes (75%).Conclusions: TTT seems to be effective for the treatment of atypical CSC in the short term. Additional studies are necessary to evaluate the long-term effectiveness and safety.Keywords: transpupillary thermotherapy, central serous chorioretinopathy, optical coherence tomography, fluorescein angiography, serous detachmen

Nectin-2 is a potential target for antibody therapy of breast and ovarian cancers

BACKGROUND: Nectin-2 is a Ca(2+)-independent cell-cell adhesion molecule that is one of the plasma membrane components of adherens junctions. However, little has been reported about the involvement of Nectin-2 in cancer. METHODS: To determine the expression of Nectin-2 in cancer tissues and cancer cell lines, we performed gene expression profile analysis, immunohistochemistry studies, and flow cytometry analysis. We also investigated the potential of this molecule as a target for antibody therapeutics to treat cancers by generating and characterizing an anti-Nectin-2 rabbit polyclonal antibody (poAb) and 256 fully human anti-Nectin-2 monoclonal antibodies (mAbs). In addition, we tested anti-Nectin-2 mAbs in several in vivo tumor growth inhibition models to investigate the primary mechanisms of action of the mAbs. RESULTS: In the present study, we found that Nectin-2 was over-expressed in clinical breast and ovarian cancer tissues by using gene expression profile analysis and immunohistochemistry studies. Nectin-2 was over-expressed in various cancer cell lines as well. Furthermore, the polyclonal antibody specific to Nectin-2 suppressed the in vitro proliferation of OV-90 ovarian cancer cells, which express endogenous Nectin-2 on the cell surface. The anti-Nectin-2 mAbs we generated were classified into 7 epitope bins. The anti-Nectin-2 mAbs demonstrated antibody-dependent cellular cytotoxicity (ADCC) and epitope bin-dependent features such as the inhibition of Nectin-2-Nectin-2 interaction, Nectin-2-Nectin-3 interaction, and in vitro cancer cell proliferation. A representative anti-Nectin-2 mAb in epitope bin VII, Y-443, showed anti-tumor effects against OV-90 cells and MDA-MB-231 breast cancer cells in mouse therapeutic models, and its main mechanism of action appeared to be ADCC. CONCLUSIONS: We observed the over-expression of Nectin-2 in breast and ovarian cancers and anti-tumor activity of anti-Nectin-2 mAbs via strong ADCC. These findings suggest that Nectin-2 is a potential target for antibody therapy against breast and ovarian cancers

Two Types of Polyp Shape Observed in the Stomach of Patients with Peutz-Jeghers Syndrome

The characteristics of gastric polyps in patients with Peutz-Jeghers (PJ) syndrome (PJS) have not been fully investigated. The objective of this study was to reveal the endoscopic and pathologic findings of gastric polyps in patients with PJS. We reviewed 11 patients with PJS treated at 6 institutions, and summarized the endo-scopic and pathologic features of their gastric polyps. The polyps were mainly classified into 2 types: (i) soli-tary or sporadic polyps > 5 mm, reddish in color with a sessile or semi-pedunculated morphology (n = 9); and (ii) multiple sessile polyps ≤ 5 mm with the same color tone as the peripheral mucosa (n = 9). Patients who underwent endoscopic mucosal resection for polyps > 5 mm were diagnosed with PJ polyps (n = 2), whereas those who underwent biopsy were diagnosed with hyperplastic polyps. Polyps ≤ 5 mm were pathologically diagnosed as fundic gland polyps or hyperplastic polyps. This study revealed that patients with PJS present with 2 types of polyps in the stomach. Endoscopic mucosal resection of polyps > 5 mm seems necessary for the pathologic diagnosis of PJ polyps

Z-360, a novel therapeutic agent for pancreatic cancer, prevents up-regulation of ephrin B1 gene expression and phosphorylation of NR2B via suppression of interleukin-1 β production in a cancer-induced pain model in mice

<p>Abstract</p> <p>Background</p> <p>Z-360 is an orally active cholecystokinin-2 (CCK2)/gastrin receptor antagonist currently under development as a therapeutic drug for pancreatic cancer. It was previously reported that Z-360 treatment in combination with gemcitabine prolonged the survival period in a lethal pancreatic cancer xenograft model in mice. In a phase Ib/IIa clinical study, Z-360 treatment displayed a trend of reduced pain in patients with advanced pancreatic cancer in combination with gemcitabine including analgesics such as opioids. Here, we investigated the mechanism of analgesic action of Z-360 in a severe cancer-induced pain model in mice, which is considered to be opioid-resistant, by examining ephrin B1 gene expression, <it>N</it>-methyl-D-aspartate receptor NR2B subunit phosphorylation, and interleukin-1β (IL-1β) production.</p> <p>Results</p> <p>In a mouse model of cancer-induced pain, ephrin B1 gene expression in dorsal root ganglia (DRGs) and the phosphorylation of NR2B in the spinal cord were induced. Z-360 treatment inhibited both ephrin B1 gene expression and the phosphorylation of NR2B. In addition, IL-1β production increased in the cancer-inoculated hind paw of mice, but could be suppressed by treatment with Z-360. Moreover, we observed that the CCK1 receptor antagonist devazepide similarly suppressed up-regulation of ephrin B1 gene expression and IL-1β production, and that the intraperitoneal injection of sulfated CCK-8 induced the production of IL-1β in the cancer-inoculated region.</p> <p>Conclusions</p> <p>We have identified a novel pain cascade, in which IL-1β production in cancer-inoculated regions induces ephrin B1 gene expression in DRGs and then ephrin B1 enhances the tyrosine phosphorylation of NR2B via Eph B receptor in the spinal cord. Notably, Z-360 relieves cancer-induced pain by preventing this pain cascade through the suppression of IL-1β production, likely via the blockade of CCK1 receptor. The pre-clinical results presented here support the analgesic action of Z-360 in pancreatic cancer patients with severe, opioid-resistant pain. Pre-clinical and clinical results have demonstrated that Z-360 combined with gemcitabine represents a promising pancreatic cancer therapy approach with characteristic analgesic effects in addition to the prolongation of survival.</p

Computed tomography findings of intersigmoid hernia

Purpose: To evaluate the computed tomography findings of intersigmoid hernias. Material and methods: Between April 2010 and March 2018, 7 patients who were surgically diagnosed with intersigmoid hernia in 3 institutions were enrolled in this study. Two radiologists evaluated imaging findings for the herniated small bowel, the distance between the occlusion point and bifurcation of the left common iliac artery, and the anatomic relationship with adjacent organs. Results: All patients were male, and their mean age (standard deviation, range) was 61.0 (13.5, 36-85) years. The mean size of the bowel loops was 5.2 (1.3, 4.0-8.3) cm in the caudal direction, 3.6 (0.8, 2.5-5.1) cm in the lateral, and 3.4 (0.6, 2.5-4.7) cm in the anterior-posterior direction. The volume was 37.9 (27.8, 15.6-103.0) cm3 approximated by an ellipse, and 24.0 (17.7, 9.9-65.6) cm3 approximated by a truncated cone. The obstruction point was located 3.6 (0.6, 2.8-4.7) cm inferior to the bifurcation of the left common iliac artery. In all cases, the small bowel ran under the point at which the inferior mesenteric vessels bifurcated to the superior rectal vessels and the sigmoid vessels and formed a sac-like appearance between the left psoas muscle and the sigmoid colon. The ureter ran dorsal to the point of the bowel stenosis, and the left gonadal vein ran outside the small bowel loops. Conclusions: All cases showed common imaging findings, which may be characteristic of men's intersigmoid hernia. In addition, the fossa's position was lower, and the size was larger than in the previous study, which may be a risk factor