Evidence Of Biological Activity Research Articles

UV-LASER adjuvant-surfactant-facilitated delivery of mobile dsRNA to tomato plant vasculature and evidence of biological activity by gene knockdown in the potato psyllid.

Double-stranded RNA (dsRNA) biopesticides are of interest for the abatement of insect vectors of pathogenic bacteria such as 'Candidatus Liberibacter', which infects both its psyllid and plant hosts. Silencing of genes essential for psyllids, or for Liberibacter, is anticipated to lead to mortality or impeded bacterial multiplication. Foliar delivery is preferred for biopesticide application; however, the cuticle impedes dsRNA penetration into the vasculature. Here, conditions were established for wounding tomato leaves using ultraviolet light amplification by stimulated emissions of radiation (UV-LASER) to promote dsRNA penetration into leaves and vasculature. UV-LASER treatment with application of select adjuvants/surfactants resulted in vascular delivery of 100-, 300- and 600-bp dsRNAs that, in general, were correlated with size. The 100-bp dsRNA required no pretreatment, whereas 300- and 600-bp dsRNAs entered the vasculature after UV-LASER treatment only and UV-LASER adjuvant/surfactant treatment, respectively. Of six adjuvant/surfactants evaluated, plant-derived oil combined with an anionic organosilicon compound performed most optimally. Localization of dsRNAs in the tomato vasculature was documented using fluorometry and fluorescence confocal microscopy. The biological activity of in planta-delivered dsRNA (200-250 bp) was determined by feeding third-instar psyllids on tomato leaves post UV-LASER adjuvant/surfactant treatment, with or without psyllid cdc42- and gelsolin dsRNAs. Gene knockdown was quantified by quantitative, real-time polymerase chain reaction with reverse transcription (RT-qPCR) amplification. At 10 days post the ingestion-access period, knockdown of cdc42 and gelsolin expression was 61% and 56%, respectively, indicating that the dsRNAs delivered to the tomato vasculature were mobile and biologically active. Results indicated that UV-LASER adjuvant/surfactant treatments facilitated the delivery of mobile, biologically active dsRNA molecules to the plant vasculature. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The significance of iron ooids from the middle Eocene of the Transylvanian Basin, Romania

The middle Eocene ironstone of the Transylvanian Basin, Romania, provides new insight into the genesis and paleoenvironmental significance of ferruginous ooids. An horizon at the base of the Căpuş Formation in north-western Romania, well known for the spectacular nummulite banks there exposed, documents a peculiar association of large foraminifera within a matrix dominated by millimetric Fe-ooids. These red-brownish Fe-ooids are well evident through the whitish colour of the carbonatic host matrix. An integrated analysis of the ooids, carried out by Environmental Scanning Electron Microscopy, Micro-X-ray Diffraction and Raman spectroscopy, allowed its detailed chemical and mineralogical characterization. Ooids are mostly composed of continuous concentric cortical layers of goethite interlayered by a few thin layers of phosphatic phases, whereas nuclei are made of ferruginous grains. No evidence of biological activity was detected in the cortex of the ooids to support a bio-mediated genesis. The occurrence of this ironstone testifies the existence of ferruginous bottom waters in the Eocene tropical/sub-tropical shallow-marine settings of the central Neotethyan Realm. The limited extension of the iron source area suggests that also local (e.g., not global) events could have been able to trigger iron ooidal deposition throughout the Phanerozoic.

Azithromycin to Prevent Recurrent Wheeze Following Severe Respiratory Syncytial Virus Bronchiolitis

BackgroundEarly-life severe respiratory syncytial virus (RSV) bronchiolitis is a risk factor for childhood asthma. Because azithromycin may attenuate airway inflammation during RSV bronchiolitis, we evaluated whether it would reduce the occurrence of post-RSV recurrent wheeze.MethodsWe prospectively enrolled 200 otherwise healthy 1- to 18-month-old children hospitalized with RSV bronchiolitis in this single-center, double-blind, placebo-controlled study and randomly assigned them to receive oral azithromycin (10 mg/kg daily for 7 days, followed by 5 mg/kg daily for 7 days) or placebo. Randomization was stratified by recent open-label antibiotic use. The primary outcome was the occurrence of recurrent wheeze, defined as a third episode of post-RSV wheeze over the following 2 to 4 years.ResultsAs an indication of the biologic activity of azithromycin, nasal wash interleukin-8 levels, at day 14 after randomization, were lower among azithromycin-treated participants (P<0.01). Despite evidence of biologic activity, azithromycin did not reduce the risk of post-RSV recurrent wheeze (47% in the azithromycin group vs. 36% in the placebo group; adjusted hazard ratio, 1.45; 95% confidence interval [CI], 0.92 to 2.29; P=0.11). Azithromycin also did not modify the risk of recurrent wheeze among participants already receiving other antibiotic treatment at the time of enrollment (hazard ratio, 0.94; 95% CI, 0.43 to 2.07). There was a potential signal among antibiotic-naïve participants who received azithromycin to have an increased risk of recurrent wheeze (hazard ratio, 1.79; 95% CI, 1.03 to 3.1).ConclusionsAzithromycin therapy for 14 days during acute severe RSV bronchiolitis did not reduce recurrent wheeze occurrence over the following 2 to 4 years. Our data suggest no benefit of azithromycin administration with the goal of preventing recurrent wheeze in later life. (Funded by the National Heart, Lung, and Blood Institute; ClinicalTrials.gov number, NCT02911935.)

Palaeoproterozoic (2.2 Ga) life on land near Medicine Bow Peak, Wyoming, U.S.A.

Tubular megafossils from the Palaeoproterozoic (ca. 2.3 Ga) Medicine Peak Quartzite of Wyoming have been regarded as possibly the earliest metazoan burrows. This interpretation has been controversial because these fossils are much older than accepted traces of metazoans. Additional similar specimens have now been found in palaeosols of the overlying Sugarloaf Quartzite (ca. 2.2 Ga), in the same area. These newly discovered fossils postdate fluvial deposition, predate metamorphic veining, and formed during oxidative, red, gypsic, soil formation. Oxidized tubular features lack regularity of width, backfills, scratches, or other complexities of metazoan trace fossils, and are more like cyanobacterial ropes, slime mold slugs, and fungal and lichen thalli of biological soil crusts. Because exact biological affinities are unknown, the fossils are assigned to palaeobotanical form taxa like many Precambrian fossils: Erythronema ramosum and E. robustum gen. et sp. nov. and Koilosolos pravus gen. et sp. nov. Such vertical, irregularly tubular fossils, which destroy prior bedding, are evidence of biological activity in Palaeoproterozoic palaeosols.

Abstract CT113: Preliminary data on a phase 1b, first-in-human study of the allosteric extracellular FGFR2 inhibitor alofanib in patients with refractory metastatic gastric cancer

Abstract Fibroblast growth factor receptor 2 (FGFR2) is associated with an unfavorable prognosis in patients with gastric cancer. Acquired mutations in FGFR2 develop resistance to multikinase inhibitors. Besides, resistance to monoclonal antibodies depends on the type of FGFR2 isoforms IIIc or IIIb expressed by cancer cells. Alofanib (RPT835) is a small molecule, allosteric inhibitor that binds to the non-active site of FGFR2 extracellular domain. RPT835GC1B (NCT04071184) is an ongoing Phase 1b open-label study evaluating the safety and preliminary efficacy of alofanib in patients with metastatic gastric adenocarcinoma pretreated with ≥ 1 previous lines of therapy. The standard dose-escalation part (design 3+3) aims to establish the maximum tolerated dose (MTD) or recommended phase 2 dose (R2PD) as a primary endpoint. The first part of the study includes a 28-day period when alofanib is administered daily intravenously for 5-days followed by a 2-day interval (rest). There are five dose levels: 50, 100, 165, 250, and 350 mg/m2. Secondary endpoints include pharmacokinetic (PK) parameters, rate of adverse events, progression-free survival (PFS), overall survival (OS), and objective response rate. All patients received alofanib until disease progression or unacceptable toxicity. As of data cutoff on December 30, 2020, 13 patients have been enrolled in the trial. Patients were predominantly male (85%), 54% had 2 and more metastatic sites, including liver metastases (54%), and were heavily pretreated (60% received previous 3-6 lines of therapy). To date, all enrolled patients have been studied for safety and have not experienced any dose-limiting toxicities (DLTs) within the 28-day DLT-assessment window. Grade 3 or higher drug related adverse events have included raised ALT/AST at 50 mg/m2, diarrhea at 165 mg/m2, and hyponatremia at 350 mg/m2. 93% of patients had any grade adverse events. Most common Grade 1-2 adverse events included fatigue, diarrhea, nausea, anemia, thrombocytopenia, increased alkaline phosphatase, and reactions immediately after intravenous injections (facial flushing, dizziness, weakness, sweating, and sinus tachycardia). Grade 1 hyperphosphatemia was founded in 25% of cases. One patient discontinued treatment due to drug related Grade 3 uncontrolled diarrhea. Alofanib has demonstrated evidence of biologic activity in 12 patients in the first 4 dose levels evaluated to date. Disease control rate was 75% (1 durable partial response (13 months) at 50 mg/m2 and 8 stable diseases at 50-250 mg/m2). After a median follow-up of 4.5 months, the median PFS and OS was not reached. In conclusion, dosing up to 350 mg/m2 of alofanib was well tolerated, DLT and MTD were not reached. The early biologic activity of alofanib in the late-line treatment of metastatic gastric cancer is encouraging. In addition to the above, any updated safety, PK, and efficacy data will be presented at the time of the AACR meeting. Citation Format: Ilya Tsimafeyeu, Galina Statsenko, Anastasia Mochalova, Natalia Trenina, Vyacheslav Kurakin, Natalia Besova, Kristina Novoselova, Natalia Abramova, Ekaterina Obarevich, Alina Kashanova, Margarita Suetina, Vladimir Moiseyenko, Mikhail Byakhov, Elena Artamonova, Liubov Vladimirova, Sergei Tjulandin. Preliminary data on a phase 1b, first-in-human study of the allosteric extracellular FGFR2 inhibitor alofanib in patients with refractory metastatic gastric cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr CT113.

Efficacy and safety of abatacept in active primary Sjögren’s syndrome: results of a phase III, randomised, placebo-controlled trial

ObjectivesTo evaluate efficacy and safety of abatacept in adults with active primary Sjögren’s syndrome (pSS) in a phase III, randomised, double-blind, placebo-controlled trial.MethodsEligible patients (moderate-to-severe pSS [2016 ACR/European League Against...

Abstract 3376: HER2-XPAT and EGFR-XPAT: Pro-drug T-cell engagers (TCEs) engineered to address on-target, off-tumor toxicity with potent efficacy in vitro and in vivo and large safety margins in NHP

Abstract Background: TCEs have proven to be effective in creating durable remissions in hematologic malignancies but have been challenging in solid tumors due to on-target, off-tumor toxicity. To circumvent the toxicity, many have tried step-up, fractionated dosing or complex formats, but these have largely been unsuccessful due to toxicity and/or enhanced immunogenicity. To address this challenge, Amunix has developed a conditionally active TCE, the XPAT or XTENylated Protease-Activated T Cell Engager, that exploits the protease activity present in tumors vs. healthy tissue, enabling expansion of the therapeutic index (TI). The core of the XPAT consists of 2 tandem scFVs targeting CD3 and a tumor antigen. Two unstructured and modular polypeptide masks (XTEN) are attached to the core that sterically reduce target engagement and extend T1/2. Protease cleavage sites at the base of these XTEN polypeptide masks are then proteolytically released in the tumor microenvironment to unleash a highly potent TCE with a short T1/2, further improving the TI. Methods: HER2-XPAT and EGFR-XPATs were each purified from E.coli expressed from a single plasmid. The activity of both the prodrugs (XPATs) and their protease-activated counterparts (PATs) were characterized for cytotoxicity in vitro and in huPBMC-transduced tumor-bearing mice. Pilot toxicity studies were conducted in NHPs (cyno). Results: Both protease-activated (PATs) forms of EGFR-XPAT and HER2-XPAT demonstrated potent in vitro T cell directed cytotoxicity against tumor cells (EC50s 1-2pM), while the masked XPAT provided 3,000-14,000-fold protection against target cell killing. Intact EGFR-XPAT demonstrated dose-dependent complete tumor responses (CRs) in HT-29 CRC xenografts, with HER2-XPAT demonstrating similar efficacy and CRs in both BT-474 and SK-OV-3 tumors. In cynos, 1 mg/kg of EGFR-XPAT was the MTD and 1.5 mg/kg exceeded the MTD, while the activated PAT was lethal at 0.132mg/kg/day by continuous infusion (with single dose 1 hour bolus of 0.066ug/kg). This indicated a near 2 log protection or more in predicted Cmax of XPAT vs PAT. A HER2-XPAT has been dose escalated up to 21 mg/kg in cynos and was well-tolerated, with the MTD still unreached. At doses starting from 2.5mg/kg of a HER2-XPAT, lymphocyte margination and laboratory abnormalities were observed, indicating evidence of biologic activity well below the tolerated 21 mg/kg dose. An estimated T1/2 of ~4 days was observed. Conclusions: EGFR-XPAT and HER2-XPAT are novel T cell engagers with protease-cleavable XTEN masks with preclinical evidence of potent anti-tumor activity and a wide margin of protection in cynos. With the prior XTEN clinical data of low immunogenicity, the XPAT TCEs provide a promising solution to overcome On-target, Off-tumor toxicity. Additional PD, safety and efficacy data will be presented. Citation Format: Fiore Cattaruzza, Ayesha Nazeer, Zachary Lange, Mikhail Hammond, Caitlin Koski, Angela Henkensiefken, Mika K. Derynck, Bryan Irving, Volker Schellenberger. HER2-XPAT and EGFR-XPAT: Pro-drug T-cell engagers (TCEs) engineered to address on-target, off-tumor toxicity with potent efficacy in vitro and in vivo and large safety margins in NHP [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3376.

The repaired durophagous scar in the shell of a lingulid brachiopod immediately after the end Ordovician mass extinction

Repaired shell injuries, widely reported mainly on mollusks, give first hand evidence of biological activity in fossil communities. Scars in fossil brachiopods from predation are rarely found, particularly after major biotic crises. A repair scar on a lingulid brachiopod Pseudolingula? sp. that lived immediately after the end Ordovician mass extinction is reported for the first time here. The specimen, cohabited with other components of Cathaysiorthis fauna, was collected from the basal Shiyang Formation (earliest Rhuddanian, Llandovery, Silurian), Jiangxi, southeast China. The scar’s near-margin position and its drape-like shape, together with the quiet middle to deeper shelf living environment and nearly infaunal lifestyle of the species, suggest that the shell damage was biotic-induced, from a failed predation, possibly by a nautiloid cephalopod. The successful repair supports previous studies showing that environmental conditions ameliorated quickly after the extinction. To analyze its repair mode, another repaired specimen of its living relative Lingula anatina collected from the Yellow Sea, China was examined here as well. Compared to the much more efficient repair for the living lingulids, the drape-like structure of the scar on the shell of Pseudolingula? sp. is more similar to the published repair specimens of Cambrian lingulate brachiopods, which may indicate the evolution of its repair mechanisms.Bing Huang [bhuang@nigpas.ac.cn], State Key Laboratory of Palaeobiology and Stratigraphy, Nanjing Institute of Geology and Palaeontology, Center for Excellence in Life and Paleoenvironment, Chinese Academy of Sciences, 39 East Beijing Road, Nanjing 210008, China; Rongyu Li [lir@brandonu.ca] Department of Geology, Brandon University, 270-18th Street, Brandon, Manitoba R7A 6A9, Canada.

Oxidative stress mediated by gyrophoric acid from the lichen Umbilicaria hirsuta affected apoptosis and stress/survival pathways in HeLa cells

BackgroundLichens produce a huge diversity of bioactive compounds with several biological effects. Gyrophoric acid (GA) is found in high concentrations in the common lichen Umbilicaria hirsuta, however evidence for biological activity was limited to anti-proliferative activity described on several cancer cell lines.MethodsWe developed and validated a new protocol for GA isolation, resulting in a high yield of highly pure GA (validated by HPLC and NMR) in an easy and time saving manner. Anti-proliferative and pro-apoptotic activity, oxygen radicals formation and stress/survival proteins activity changes was study by flow cytometry.ResultsThe highly purified GA showed anti-proliferative activity against HeLa (human cervix carcinoma) and other tumor cells. Moreover, GA threated cells showed a significant increase in caspase-3 activation followed by PARP cleavage, PS externalization and cell cycle changes mediated by oxidative stress. Production of oxygen radicals led to DNA damage and changes in stress/survival pathways activation.ConclusionsGA treatment on HeLa cells clearly indicates ROS production and apoptosis as form of occurred cell death. Moreover, DNA damage and changing activity of stress/survival proteins as p38MAPK, Erk1/2 and Akt mediated by GA treatment confirm pro-apoptotic potential. The pharmacological potential of U. hirsuta derived GA is discussed.

Abstract P6-21-03: Phase I trial of intratumoral (IT) administration of a NIS-expressing derivative manufactured from a genetically engineered strain of measles virus (MV)

Abstract Background: The live attenuated non-pathogenic Edmonston MV vaccine strain has advantages as an oncolytic platform given its tumor specificity, potent bystander effect, and ability to be engineered and retargeted. MV-NIS expresses the human thyroidal sodium-iodide symporter (NIS) and is selectively oncolytic, entering tumor cells through CD46 (overexpressed on many cancers, including breast cancer of all subtypes) and Nectin-4. NIS expression in MV-NIS infected cells permits noninvasive monitoring of virus spread by SPECT-CT imaging of Tc-99m pertechnetate or I-123 uptake. Methods: NCT01846091 is a standard 3+3 phase I trial of a single IT administration of MV-NIS in pts with recurrent/metastatic squamous cell carcinoma of the head and neck (SCCHN) or metastatic breast cancer (MBC). Primary objectives are (a) safety and tolerability and (b) maximally tolerated single dose. The secondary clinical objective is to preliminarily assess antitumor efficacy at and away from the MV injection site. Key eligibility criteria were: absence of standard therapy with life prolonging intent; at least one lesion &amp;gt;1 cm amenable to percutaneous injection; and no impending visceral crisis. MV-NIS was administered on D1 with mandatory SPECT-CT at baseline (BL) and on D3&amp;D8; repeat SPECT-CT on D15&amp;D21 if the prior result was positive; mandatory tumor biopsies on D3&amp;D21; optional tumor biopsies on D8&amp;D15; assessments for viremia and viral shedding at BL and on D3,D8,D15,D21; and standard imaging for restaging at BL,D21,W6,W12. Results: Accrual completed with 12 evaluable pts (6 SCCHN and 6 MBC) at 3 dose levels (108, 3x108, 109 TCID50). The MBC group included 5 HR+/HER2- pts and 1 pt with mixed HR+/HER2- and HR+/HER2+ disease. 5 pts had evidence of disease progression prior to study participation. No dose limiting toxicities were observed among the MBC pts; AEs possibly related to MV-NIS in this group were gr2 fatigue, gr1 flu-like illness, gr2 lymphopenia (all n=1). No SCCHN responses were observed. Best response for the MBC pts was: stable disease (SD) &amp;gt;6 wks, n=4; clinical response, n=1; progression, n=1. One MBC pt with SD for 12 wks had positive SPECT/CT imaging at and away from the injection site on D3&amp;D8 and was the only pt seronegative for measles IgG antibodies prior to MV-NIS exposure. The MBC pt who responded after initial MV-NIS exposure was the only pt with low viral RNA in blood (D3); she received additional doses at W9&amp;W13 without toxicity through an expanded access protocol exemption and had disease progression by W19. No viral shedding was detected from mouth rinse or urine in any pt. MV was detected in tumor samples from all pts treated at the highest dose level. Additional blood and tissue analyses are in progress. Conclusion: These results demonstrate the safety of IT MV-NIS administration, provide early evidence of biologic activity in MBC, and support the possibility of viral replication in tumors remote from the IT injection site. A MV strain encoding the immunomodulatory neutrophil activating protein transgene has been constructed (MV-s-NAP) with preclinical evidence of improved antitumor activity and immunogenicity. The phase I MV-s-NAP trial will start recruitment in Fall 2018. Citation Format: Liu MC, Peng K-W, Federspiel MJ, Russell SJ, Brunton BA, Zhou Y, Packiriswamy N, Hubbard JM, Loprinzi CL, Peethambaram PP, Ruddy KJ, Allred JB, Galanis E, Okuno SH. Phase I trial of intratumoral (IT) administration of a NIS-expressing derivative manufactured from a genetically engineered strain of measles virus (MV) [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P6-21-03.

Abstract IA24: Translating fundamental immunobiology into adoptive T-cell therapy for ovarian cancer

Abstract In nearly half of all high-grade serous cancers, tumor-infiltrating T lymphocytes (TILs) invade and accumulate in tumor epithelial islets. These immunologically “hot” cancers are prognostic for patient survival and provide a strong rationale for immune-based treatment strategies that mobilize and enhance endogenous immune responses. In other cancers, such as melanoma and HPV-associated cervical cancer, TILs with autologous tumor reactivity can be isolated from resected tumors and expanded ex vivo for reinfusion to the patient in a process called adoptive immunotherapy. Adoptive immunotherapy using TILs in patients with these cancers frequently results in durable complete responses. In ovarian cancer patients, responses to TIL therapy have been less dramatic due in part to technical difficulties in identifying and expanding the tumor-reactive subset of TILs. In an effort to improve TIL therapy, we identified a surface biomarker of naturally occurring tumor reactive TILs in ovarian cancer and showed in preclinical models systems that these cells mediate enhanced TIL killing of autologous tumor cells ex vivo and in vivo, paving the way for more effective TIL therapy. Mass cytometry analysis revealed a complex network of immune inhibitory molecules expressed by tumor-reactive TILs that act in unison to dampen antitumor immunity, and for which clinical-grade blocking antibodies exist, rationalizing the use of TIL therapy in combination with immune checkpoint inhibitors for ovarian cancer. Facilitating adoptive T-cell therapy in the subset of patients with immunologically “cold” ovarian cancers relies upon the de novo induction of T cell antitumor reactivity. One approach relies on the stimulation of nonreactive endogenous immune cells with tumor antigens. Use of lysates generated from the patient’s own tumor provides the opportunity to vaccinate against shared antigens as well as patient-specific mutational antigens. In this prime-and-boost approach, a patient can first be vaccinated against patient-specific antigens to induce antitumor immunity, and the vaccine-primed T cells then harvested and expanded to high numbers outside of the body before reinfusion into the patient. The feasibility and safety of this approach has now been established in ovarian cancer with evidence of biologic activity. In the era of synthetic biology, “off-the-shelf” tumor-reactive T cells can now be generated de novo through the use of gene engineering. Using advanced gene-transfer systems and sophisticated cell-cultivation technologies, patients’ T cells are endowed with antigen-specificity and a capacity for enhanced antigen-dependent proliferation, activity, and survival, based on principles of fundamental immunology. In this way, engineered T cells can overcome issues of HLA loss by tumor cells as well as T-cell exhaustion. In preclinical xenograft models, gene-modified T cells exert antigen-specific killing of human ovarian cancer. Gene-engineered T cells are the subject of our ongoing clinical trials and pave a new route toward adoptive immunotherapy, which renders ovarian cancers immunologically “hot.” Citation Format: Daniel J. Powell, Jr. Translating fundamental immunobiology into adoptive T-cell therapy for ovarian cancer. [abstract]. In: Proceedings of the AACR Conference: Addressing Critical Questions in Ovarian Cancer Research and Treatment; Oct 1-4, 2017; Pittsburgh, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(15_Suppl):Abstract nr IA24.

Abstract CT109: HITM-SIR: Phase Ib trial of CAR-T hepatic artery infusions and selective internal radiation therapy for liver metastases

Abstract OBJECTIVES: There are no effective treatment options for patients with unresectable CEA+ liver metastases (LM) from gastrointestinal adenocarcinoma refractory to conventional systemic therapy. In the previous Hepatic Immunotherapy for Metastases (HITM) phase I study we demonstrated the safety and biologic activity of anti-CEA CAR-T cell hepatic artery infusions (HAI). HITM-SIR was a single arm phase Ib trial testing anti-CEA CAR-T HAI followed by selective internal radiation therapy (SIRT) in patients with refractory CEA+ LM. METHODOLOGY: We enrolled 8 patients with unresectable, chemotherapy refractory CEA+ LM and 6/8 completed the study. Two patients were withdrawn for disease progression prior to CAR-T infusion and biliary obstruction due to centrally located disease. Limited extrahepatic disease (EHD) was permitted. Subjects received 3 HAI of anti-CEA CAR-T cells (1e10 cells per dose) along with low dose continuous IL-2 infusions (50,000 IU/kg/day). SIRT was administered in standard fashion 2 weeks following the 3rd CAR-T HAI. Primary objective was to establish safety of the CAR-T/SIRT combination. Secondary objectives included response assessed by modified RECIST (mRECIST), immune-related response criteria (irRC), and tumor marker kinetics. RESULTS: The mean age for enrolled subjects was 54.6 years (39-61) with 3 women and 5 men. Histologies (completed patients): 2 colon, 2 rectal, and 2 pancreas. This heavily pre-treated, advanced disease group of patients received an average of 2.3 lines of prior chemotherapy, 3/6 had &amp;gt;10 LM, and the average largest LM size was 7.3 cm. The average transduction efficiency as measured by CAR expression was 60.4%, with 90.9% viability, and an average production time for &amp;gt;3e10 cells of 13.8 days. There were no grade (G) 4/5 events related to the CAR-T, SIRT, or combination. Toxicities included G 1/2 liver function test elevations (n=5/6), fever (n=5/6), hypereosinophilia (n=2/6), and edema (n=2/6). G3 toxicities included colitis (n=2/6), fever (n=2/6), and edema (n=2/6). One patient experienced a hypertensive crisis during a single CAR-T infusion but tolerated 2 infusions without incident. All colitis episodes resolved with IL-2 dose reductions. Post-SIRT serum CEA decreases were noted in 2/6 patients (-40% and -71%) with CA19-9 decreases in 2/5 detectable patients (-31% and -32%). When considering all on-study time points, 5/6 patients had CEA responses (mean decrease 59.7%) and 4/5 patients expressing CA 19-9 decreases (mean 59.6%). At completion of the study, 3/6 patients had stable disease (SD) in the liver by mRECIST and irRC, and 3/6 SD overall by irRC. Target liver lesion decreases in size among patients with SD ranged from 6-28%. Regression of extrahepatic tumors or abscopal effects were noted in 2/6 at lung and bone sites. One patient remains without evidence of viable liver disease at 10.8 months follow-up. The median overall survival time for all patients is 6.9 months (range 3.8-10.8+). CONCLUSIONS: Following this phase Ib trial, the recommended phase 2 dose for anti-CEA CAR-T HITM infusions is 1010 cells with or without SIRT. The favorable safety profile and evidence of biologic activity indicate that CAR-T HITM infusions should be further studied in a phase 2 trial. Clinical trial information: NCT02416466. Citation Format: Steven C. Katz, Ethan Prince, Marissa Cunetta, Prajna Guha, Ashley Moody, Vincent Armenio, Li J. Wang, N. Joseph Espat, Richard P. Junghans. HITM-SIR: Phase Ib trial of CAR-T hepatic artery infusions and selective internal radiation therapy for liver metastases [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr CT109. doi:10.1158/1538-7445.AM2017-CT109

Dose escalation results from a first-in-human, phase 1 study of the glucocorticoid-induced TNF receptor-related protein (GITR) agonist AMG 228 in patients (Pts) with advanced solid tumors.

2521 Background: AMG 228 is an agonistic human IgG1 monoclonal antibody that binds to GITR (CD357), a TNFSFR costimulatory molecule expressed by effector/regulatory T cells. Dose escalation of this open label, first in human, phase 1 study evaluated the safety, pharmacokinetics (PK), pharmacodynamics, and maximum tolerated dose (MTD) and recommended phase 2 dose of AMG 228 in pts with advanced solid tumors. Methods: Pts with refractory advanced colorectal cancer (n = 13), squamous cell carcinoma of head and neck (n = 10), non–small cell lung cancer (n = 2), urothelial transitional cell carcinoma (n = 4), and melanoma (n = 1) received AMG 228 IV Q3W. Dose escalation was in two stages: single-pt cohorts until AMG 228-related grade &gt; 2 adverse events (AEs), efficacy, or 90 mg dose reached (4 cohorts: 3, 9, 30, and 90 mg), followed by rolling six design (n = 2 to 6) until MTD or highest planned dose of 1200 mg reached (5 cohorts: 180, 360, 600, 900, and 1200 mg). Primary endpoints included incidence of dose-limiting toxicities (DLTs) and AEs and PK. Additional endpoints were objective response (RECIST 1.1) and evidence of biological activity. Results: In total, 30 pts (median age 63 y) were treated (3, 9, 30, and 90 mg, n = 1; 180 mg, n = 6; 360 mg, n = 4; 600 mg, n = 6; 900 mg, n = 4; 1200 mg, n = 6). Twenty-seven (90%) pts had treatment-emergent AEs; the most common were hypophosphatemia (23%), fatigue (23%), anemia (23%), nausea (20%), and pyrexia (20%). No DLTs occurred; the MTD was not reached. AMG 228 exposure was dose-related, with PK profiles at low doses (3 to 90 mg) consistent with target mediated drug disposition; doses &gt; 360 mg achieved serum levels needed for 95% receptor occupancy on activated PBMCs. No evidence of T cell activation was observed despite complete target coverage in both tumor and peripheral blood. Among 29 evaluable pts, none had an objective response. Conclusions: In this population of pts with advanced solid tumors, AMG 228 Q3W was tolerable up to the highest tested dose (1200 mg), showing favorable PK. However, no clinical or immunological activity was observed in this limited number of pts. Clinical trial information: NCT02437916.

Abstract B72: Long-term survival and outcome update on patients (pts) with recurrent ovarian cancer who received Ipilimumab post GVAX vaccine.

Abstract Immunotherapeutic approaches are amongst the most exciting new therapies for cancer, and trials of immunotherapy agents to treat ovarian cancer are underway. We report on long-term outcomes of patients with advanced and recurrent ovarian cancer treated with single agent ipilimumab, a fully humanized IgG1 monoclonal antibody against CTLA-4, following receipt of GVAX vaccine, which was a lethally irradiated, autologous ovarian cancer cells engineered by adenoviral gene transfer to secrete granulocyte macrophage colony stimulating factor (GM-CSF). The objectives of this study were to determine the safety of ipilimumab, identify preliminary evidence of biologic activity and efficacy of ipilimumab post GVAX, and, as an exploratory objective, to correlate clinical benefit with the presence of antibodies to NY-ESO and p53. Results: Eleven pts who initially were treated with GVAX received ipilimumab (3 mg/kg) IV every 2 months on this study. These 11 patients had a median age of 61 years (range 38-82 years of age) at the time on enrollment and all had measureable cancer by RECIST 1.0. Eight pts had platinum resistant cancer and 3 had platinum sensitive cancer. Histologies enrolled were high grade serous (8 pts), clear cell (1pt), poorly differentiated (1 pt), and low grade serous that had evolved to high grade (1 pt). Patients were allowed to have had intervening treatment between initial GVAX vaccine and start of ipilimumab. Of the 11 pts, the median interval between GVAX and ipilimumab was 3 months, ranging from 1 month to 38 months. Overall, ipilimumab at 3 mg/kg was well tolerated; rash was grade 1 (63%), grade 2 (9%), grade 3 (9%). Diarrhea was 18% grade 2 and 18% grade 3. Other constitutional symptoms such as fatigue were 18% grade 1 and 9% grade 3. Of the 11 pts, 6 pts had disease progression by RECIST after 1 infusion of ipilimumab with overall survival post-ipilimumab ranging from 3 to 35 months; 2 of these pts had platinum sensitive recurrence. Of the 5 pts who either experienced SD (4 pts) and PR (1 pt), the number of ipilimumab infusions were 1, 2 (2 pts), 4, and 25 infusions with survival ranging from 23 to 104 months post-ipilimumab. One patient (OV65) who received 25 ipilimumab infusions from 2003 through 2011 lived for 104 months post-initiation of ipilimumab and had a PR to ipilimumab at 95 months; she had high grade serous platinum resistant cancer with spread to liver/omentum/lymph nodes, had present NY-ESO Ab's, and had a 1 month interval between GVAX and the initiation of ipilimumab infusions. OV65 received no further chemotherapy treatment after ipilimumab was started though she had evidence of cancer present during her ipilimumab treatment course; her eventual death was not cancer related. Conclusions: CTLA-4 blockade with ipilimumab can induce long-term disease control and has evidence of activity in ovarian cancer. The role of prior GVAX vaccination is not clear. Citation Format: Ursula Anne Matulonis, Stephen Hodi, Glenn Dranoff, Susana Campos, Richard Penson, Robert Soiffer, Marcus Butler. Long-term survival and outcome update on patients (pts) with recurrent ovarian cancer who received Ipilimumab post GVAX vaccine. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: Exploiting Vulnerabilities; Oct 17-20, 2015; Orlando, FL. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(2 Suppl):Abstract nr B72.

Efficacy and safety of tabalumab, an anti-BAFF monoclonal antibody, in patients with moderate-to-severe rheumatoid arthritis and inadequate response to TNF inhibitors: results of a randomised, double-blind, placebo-controlled, phase 3 study

BackgroundTabalumab is a human monoclonal antibody that neutralises B-cell activating factor.ObjectivesTo evaluate tabalumab efficacy and safety in patients with rheumatoid arthritis (RA).MethodsThis phase 3, randomised, double-blind, placebo-controlled study evaluated 456...

Advances in the design and development of oncolytic measles viruses

A successful oncolytic virus is one that selectively propagates and destroys cancerous tissue without causing excessive damage to the normal surrounding tissue. Oncolytic measles virus (MV) is one such virus that exhibits this characteristic and thus has rapidly emerged as a potentially useful anticancer modality. Derivatives of the Edmonston MV vaccine strain possess a remarkable safety record in humans. Promising results in preclinical animal models and evidence of biological activity in early phase trials contribute to the enthusiasm. Genetic modifications have enabled MV to evolve from a vaccine agent to a potential anticancer therapy. Specifically, alterations of the MV genome have led to improved tumor selectivity and delivery, therapeutic potency, and immune system modulation. In this article, we will review the advancements that have been made in the design and development of MV that have led to its use as a cancer therapy. In addition, we will discuss the evidence supporting its use, as well as the challenges associated with MV as a potential cancer therapeutic.

Efficacy and safety of tabalumab, an anti-B-cell-activating factor monoclonal antibody, in patients with rheumatoid arthritis who had an inadequate response to methotrexate therapy: results from a phase III multicentre, randomised, double-blind study

ObjectivesRandomised, double-blind, placebo-controlled study to evaluate efficacy and safety of tabalumab in patients with rheumatoid arthritis (RA) with inadequate responses to methotrexate (MTX-IR).Methods1041 patients with moderate–severe RA despite ongoing MTX...

MORPHOLOGICAL AND MICROMORPHOLOGICAL CHANGES IN THE STRUCTURE OF A RHODIC HAPLUDOX AS A RESULT OF AGRICULTURAL MANAGEMENT

In evaluation of soil quality for agricultural use, soil structure is one of the most important properties, which is influenced not only by climate, biological activity, and management practices but also by mechanical and physico-chemical forces acting in the soil. The purpose of this study was to evaluate the influence of conventional agricultural management on the structure and microstructure of a Latossolo Vermelho distroférrico típico (Rhodic Hapludox) in an experimental area planted to maize. Soil morphology was described using the crop profile method by identifying the distinct structural volumes called Morphologically Homogeneous Units (MHUs). For comparison, we also described a profile in an adjacent area without agricultural use and under natural regrowth referred to as Memory. We took undisturbed samples from the main MHUs so as to form thin sections and blocks of soil for micromorphological and micromorphometrical analyses. Results from the application of the crop profile method showed the occurrence of the following structural types: loose (L), fragmented (F) and continuous (C) in both profiles analyzed. In the Memory soil profile, the fragmented structures were classified as Fptμ∆+tf and Fmt∆μ, whose micromorphology shows an enaulic-porphyric (porous) relative distribution with a great deal of biological activity as indicated by the presence of vughs and channels. Lower down, from 0.20 to 0.35 m, there was a continuous soil volume (sub-type C∆μ), with a subangular block microstructure and an enaulic-porphyric relative distribution, though in this case more compact and with aggregate coalescence and less biological activity. The micromorphometrical study of the soil of the Memory Plot showed the predominance of complex pores in NAM (15.03 %), Fmt∆μ (11.72 %), and Fptμ∆+tf (7.73 %), and rounded pores in C∆μ (8.21 %). In the soil under conventional agricultural management, we observed fragmented structures similar to the Memory Plot from 0.02 to 0.20 m, followed by a volume with a compact continuous structure (C∆μ), without visible porosity and with few roots. In the MHUs under conventional management, reduction in the packing pores (40 %) was observed, mainly in the continuous units (C). The microstructure had well-defined blocks, with the occurrence of planar pores and less evidence of biological activity. In conclusion, the morphological and micromorphological analyses of the soil profiles studied offered complementary information regarding soil structural quality, especially concerning the changes in pore types as result of mechanical stress undergone by the soil.

FRI0326 Efficacy and Safety of Tabalumab, an Anti-B Cell Activating Factor Monoclonal Antibody, in Patients with Rheumatoid Arthritis Who HAD an Inadequate Response to Methotrexate Therapy: Results from A Phase 3 Multicenter, Randomized, Double-Blind Study

BackgroundTabalumab is a monoclonal antibody that neutralizes membrane-bound and soluble B cell activating factor (BAFF).ObjectivesEvaluate efficacy and safety of tabalumab, in RA pts who had an inadequate response to methotrexate...

AB0438 Efficacy and Safety of Tabalumab, an Anti-B Cell Activating Factor Monoclonal Antibody, in Patients with Rheumatoid Arthritis WHO HAD an Inadequate Response to Tnf-Alpha Inhibitors: Results from A Phase 3 Multicenter, Randomized, Double-Blind Study

BackgroundTabalumab is a monoclonal antibody that neutralizes membrane-bound and soluble B cell activating factor (BAFF).ObjectivesEvaluate efficacy and safety of tabalumab, in RA pts who had an inadequate response to ≥1...