Search

We explored patient, caregiver, and provider recommendations for development of a tool kit to implement enhanced recovery protocols (ERPs) for pediatric patients undergoing gastrointestinal surgery. ERPs are widely used for adults to decrease hospital length of stay, hospital costs, and complications while hastening patient recovery after surgery. With limited data available for ERPs among pediatric populations informed modification of adult ERPs is needed to facilitate successful implementation for pediatric surgery.Using a qualitative research design, semistructured interviews were conducted with hospital-based teams including surgeons, anesthesiologists, gastroenterologists, nursing, and physician assistants. Four in-person focus groups were held at two pediatric hospitals with patients and caregivers. Codes were developed and applied to interview and focus groups transcripts for structural content analysis. Thematic analysis guided by the Active Implementation Framework, included recommendations that informed ERP implementation tool kit development.Key components of the ERP tool kit included the need for a structured and systematic approach, leadership support from key champions, and buy-in from surgical partners and hospital management. Providers identified the need for multimodal educational materials on ERP elements for staff and patients; use of uniform checklists, care sets and an electronic repository to collect outcome data for quality assurance assessment. Patients and caregivers endorsed expansion of the team to include child-life specialists, nutritionists, and patient-parent supporters to help navigate the surgical experience.This study is the first to leverage key input from patients, caregivers, and providers to identify practical components for an ERP implementation tool kit for children undergoing gastrointestinal surgery.

Supplementary Table S1-S3 Legends and Supplementary Figure S1-S6 Legends. Legends for supplementary tables S1-S3 and supplementary figures S1-S6 which support data in the manuscript, but could not be included due to space constraints.

Purpose: Histone deacetylase inhibitors (HDACi) have recently emerged as efficacious therapies that target epigenetic mechanisms in hematologic malignancies. One such hematologic malignancy, B-cell acute lymphoblastic leukemia (B-ALL), may be highly dependent on epigenetic regulation for leukemia development and maintenance, and thus sensitive to small-molecule inhibitors that target epigenetic mechanisms.Experimental Design: A panel of B-ALL cell lines was tested for sensitivity to HDACi with varying isoform sensitivity. Isoform-specific shRNAs were used as further validation of HDACs as relevant therapeutic targets in B-ALL. Mouse xenografts of B-cell malignancy–derived cell lines and a pediatric B-ALL were used to demonstrate pharmacologic efficacy.Results: Nonselective HDAC inhibitors were cytotoxic to a panel of B-ALL cell lines as well as to xenografted human leukemia patient samples. Assessment of isoform-specific HDACi indicated that targeting HDAC1-3 with class I HDAC-specific inhibitors was sufficient to inhibit growth of B-ALL cell lines. Furthermore, shRNA-mediated knockdown of HDAC1 or HDAC2 resulted in growth inhibition in these cells. We then assessed a compound that specifically inhibits only HDAC1 and HDAC2. This compound suppressed growth and induced apoptosis in B-ALL cell lines in vitro and in vivo, whereas it was far less effective against other B-cell–derived malignancies.Conclusions: Here, we show that HDAC inhibitors are a potential therapeutic option for B-ALL, and that a more specific inhibitor of HDAC1 and HDAC2 could be therapeutically useful for patients with B-ALL. Clin Cancer Res; 21(10); 2348–58. ©2015 AACR.

Supplementary Tables S1-S3 and Supplementary Figures S1-S6. Table S1 - EC50 values for B-cell malignancies treated with HDAC inhibitors of varying isoform selectivity Table S2 - EC50 values for B-ALL cell line treated with a panel of HDAC inhibitors Table S3 - statistics for cell growth assay in Figure 3 Figure S1 - In vitro survival and PD for B-ALL cells treated with LAQ824 Figure S2 - Western analyses of B-ALL cells treated with MS275 Figure S3 - Western blots, cell growth and cell cycle analysis for B-ALL cells transduced with HDAC1,2, or 3 specific shRNA Figure S4 - Western analyses, cell growth and survival of B-ALL cell treated with Merck60 Figure S5 - Survival and western blots for B-ALL cells treated with HDACi with an apoptosis inhibitor. Figure S6 - Western blots to show DNA damage caused by HDACi or doxorubicin in Merck60 insensitive multiple myeloma cell lines.

Background Natural killer (NK) cells are highly effective and fast-acting cytolytic cells capable of eradicating target cells with limited adverse effects such as cytokine release syndrome (CRS) or graft-versus-host disease. Chimeric antigen receptors (CARs)-engineered NK cells have been recently used against leukemia with encouraging clinical outcomes.1 The surface antigen CD19, expressed by B-lymphoblasts, represents an ideal CAR target against B cell acute lymphoblastic leukemia (B-ALL). We developed a highly potent CD19 -directed CAR NK cell therapy, NKX019, with an extended in vivo half-life aimed at killing CD19-expressing target. Methods NK cells isolated from healthy PBMCs were expanded in the presence of NKSTIM cells, IL-2, IL-12, IL-18 and transduced with both a CD19-targeted CAR construct and a membrane-bound form of IL-15 (mbIL-15). Control (non-engineered) NK cells were produced in parallel. Cytotoxic activity of NKX019 against CD19+ B-ALL cell line (REH), pre-B ALL cell line (Nalm-6), allogeneic PBMCs was assessed using Incucyte® or flow cytometry. NSG mice bearing either Nalm-6.fluc (Nalm6) or REH.fluc (REH) tumor received different concentrations of NKX019 or control NK cells. In-life analysis of tumor-bearing and naive NSG mice include: 1) bioluminescence imaging, 2) clinical observations, 3) serum cytokines and 4) CAR+ NK cell persistency. Results NKX019 showed enhanced cytolytic activity against REH and Nalm-6 tumor cells compared to control NK cells and CAR19+ T cells. The superiority of NKX019 over CAR19+ T cells was more pronounced at the earlier time point (24 hours) with near identical calculated EC50 observed at 72 hours for both cell types. Increased cytolytic activity of NKX019 was limited to CD19+ cells in bulk PBMCs. Consistent with our in vitro observations, NKX019 controlled Nalm-6 and REH tumor growth in doses as low as 2 × 106 cells/kg for up to 30 days with no apparent increase in cytokines commonly associated with CRS. Increased Nalm-6 tumor growth coincided with an apparent decrease in measurable NKX019 in the periphery. In tumor-naive NSG mice, NKX019 was detectable in the blood for up to 9 weeks post-infusion consistent with its extended half-life. Conclusions NKX019 expresses mbIL-15 and is produced in the presence of IL-12 and IL-18, resulting in enhanced in vitro expansion and longer in vivo half-life than non-engineered NK cells. NKX019 also exhibited advantages compared to CAR19+ T cells including faster cytotoxic kinetics and limited production of cytokines associated with CRS. A first-in-human trial of NKX019 in B cell malignancies is planned for 2021. Ethics Approval The animal procedures described in this abstract were conducted in accordance with Explora BioLabs Animal Care and Use Protocol approved by Explora BioLabs Institutional Animal Care and Use Committee. Reference Liu, et al. 2020 NEJM

Multiple myeloma (MM) is a progressive hematological cancer with a 5-year survival rate of 53% (1). Novel therapeutic strategies are being developed to target specific MM surface antigens. Yet, changes in antigen expression through MM progression are poorly understood in the clinic and have not been well characterized in preclinical models. Here, we were interested in understanding how BCMA, CD38, CD138 and HLA-DR surface expression patterns may be affected as MM progresses in preclinical models. To do so, ARH-77 and RPMI-8226 cells were inoculated subcutaneously while MM.1S, MM.1R, U266 and NCI-H929 cells were dosed intravenously into NSG mice. Tumor xenograft or bone marrow, whole blood and spleen were processed for flow cytometry analysis. Antigen expression was measured at different stages of tumor progression as well as in vitro. Soluble BCMA levels were assessed in the mouse serum by ELISA. ARH-77 and RPMI-8226 subcutaneous xenografts show preferential growth when inoculated in PBS and 50% growth factor-reduced Matrigel, respectively. BCMA expression was reduced in both ARH-77 and RPMI-8226 xenografts compared to in vitro culture. CD38 and HLA-DR expression increased as ARH-77/PBS and RPMI-8226/Matrigel tumor volume progressed. All 4 antigens were detected in the bone marrow from MM.1S, MM.1R and U266-bearing mice, as early as 5-6 weeks post intravenous injection. In contrast, antigens were not detected in any tissue from mice injected with NCI-H929, likely indicating failed engraftment. At 5 weeks, antigens were detected in the whole blood and spleen in MM.1S-bearing mice only, suggesting organ invasion and a more advanced disease-stage compared to MM.1R or U266. Experiments are being carried out to test antigen expression in tissue at late-stage tumor progression as well as serum BCMA levels in the intravenous MM models. These data highlight antigen expression differences in MM cells when analyzed in mouse tissue compared to in vitro culture. Like the widely variable expression observed between patients (2, 3), BCMA and CD138 were differentially expressed in the mouse bone marrow between models. Our observations suggest that commonly targeted antigens in MM vary kinetically in vivo and can be measured by flow cytometry. The present findings also support the use of RPMI-8226 and MM.1S cell lines when screening for antigen-specific immunotherapies and combinatorial studies for MM treatment. References: 1.Howlader et al. https://seer.cancer.gov/csr/1975_2017 2.Brudno et al. J Clin Oncol. 2018 3.Kawano et al. Int J Oncol. 2012 Citation Format: Nadege Morisot, Julian Tam, Nicole Dailey, Tina Davis, Jacinda Chen, Janeen Islar, Luxuan Buren, Nitin Patel, Sasha Lazetic, Ivan Chan, James B. Trager, Joanne B. Tan. Surveying surface antigen expression in multiple myeloma preclinical models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6004.

Identification of a patient homozygous for familial defective apolipoprotein B-100(FDB) and successful treatment with PCSK9 inhibition.

The heart exhibits the highest basal oxygen (O2) consumption per tissue mass of any organ in the body and is uniquely dependent on aerobic metabolism to sustain contractile function. During acute hypoxic states, the body responds with a compensatory increase in cardiac output that further increases myocardial O2 demand, predisposing the heart to ischemic stress and myocardial dysfunction. Here, we test the utility of a novel engineered protein derived from the heme-based nitric oxide (NO)/oxygen (H-NOX) family of bacterial proteins as an O2 delivery biotherapeutic (Omniox-cardiovascular [OMX-CV]) for the hypoxic myocardium. Because of their unique binding characteristics, H-NOX-based variants effectively deliver O2 to hypoxic tissues, but not those at physiologic O2 tension. Additionally, H-NOX-based variants exhibit tunable binding that is specific for O2 with subphysiologic reactivity towards NO, circumventing a significant toxicity exhibited by hemoglobin (Hb)-based O2 carriers (HBOCs). Juvenile lambs were sedated, mechanically ventilated, and instrumented to measure cardiovascular parameters. Biventricular admittance catheters were inserted to perform pressure-volume (PV) analyses. Systemic hypoxia was induced by ventilation with 10% O2. Following 15 minutes of hypoxia, the lambs were treated with OMX-CV (200 mg/kg IV) or vehicle. Acute hypoxia induced significant increases in heart rate (HR), pulmonary blood flow (PBF), and pulmonary vascular resistance (PVR) (p0.05). At 1 hour, vehicle-treated lambs exhibited severe hypoxia and a significant decrease in biventricular contractile function. However, in OMX-CV-treated animals, myocardial oxygenation was improved without negatively impacting systemic or PVR, and both right ventricle (RV) and left ventricle (LV) contractile function were maintained at pre-hypoxic baseline levels. These data suggest that OMX-CV is a promising and safe O2 delivery biotherapeutic for the preservation of myocardial contractility in the setting of acute hypoxia.

Rationale Accumulation and activation of mast cells and eosinophils have been implicated in the pathogenesis of several chronic inflammatory gastrointestinal (GI) diseases, including eosinophilic gastrointestinal diseases (EGIDs) and inflammatory bowel disease (IBD). Despite the strong association of mast cell and eosinophil numbers and activation with the pathogenesis of IBD, no further characterization of these cells has been performed. Current treatment options for IBD include aminosalicylates, antibiotics, immunomodulators, biologic agents and small molecules. These therapies are only moderately effective. A significant proportion of patients fail to respond, do not fully respond, or lose response over time. Therefore, there is significant need for more selective and effective therapy options. Siglec-8 is an inhibitory receptor selectively expressed on human eosinophils and mast cells and represents a novel target for the treatment of IBD with the anti-Siglec-8 mAb, antolimab (AK002). We aimed to quantify and evaluate the activation state of mast cells and eosinophils in colonic tissue from IBD or non-diseased patients. In addition, we quantified the production of TNFa from human colon tissue mast cells and evaluated the inhibitory activity of antolimab (AK002) on these cells. Methods Single-cell suspensions were prepared by enzymatic digestion of fresh colonic biopsies from patients clinically diagnosed with IBD (n=29) or non-diseased control tissues (n=16). Multi-color flow cytometry was performed to identify major immune cell populations and evaluate the activation state of mast cells and eosinophils. Mast cells were FACS-sorted from human colon tissue to evaluate cytokine production and inhibitory activity of antolimab. Results The percentage of mast cells and the expression of the mast cell degranulation marker CD107a were significantly increased in ulcerative colitis (UC) patient biopsy tissue compared to Crohn’s disease (CD) and non-diseased colonic tissue (Figure 1A and B). Furthermore, FACS-sorted mast cells from human colon tissue produced significant quantities of TNFa that was reduced after ex vivo antolimab treatment. Colonic tissue eosinophils were also elevated in a subset of UC and CD patient biopsies, and all UC and CD tissue eosinophils displayed increased expression of the activation marker CD11b compared to control colonic tissue. Conclusions Mast cells and eosinophils may play a significant role in driving the pathogenesis of ulcerative colitis through the production of inflammatory mediators. The high expression of Siglec-8 and the inhibitory activity against mast cells suggests that antibodies that target this receptor, such as antolimab (AK002) represent a potential novel approach for the treatment of IBD.

BACKGROUND: Intratumoral hypoxia is associated with resistance to chemo- and radio-therapies and poor patient outcomes. In addition, hypoxia promotes the immune escape of tumors by altering the recruitment and function of innate and adaptive immune effector and suppressor cells. Therefore, reversing tumor hypoxia to create an immunopermissive microenvironment may improve anti-tumor response, and combined with immunotherapy approaches such as checkpoint inhibitors (CPI) may increase therapeutic efficacy. OMX, an anti-cancer therapy designed to reverse tumor hypoxia, efficiently accumulates in orthotopic rodent GB and spontaneous canine brain tumors, reduces tumor hypoxia and enhances immunotherapeutic efficacy. METHODS: We used in vivo bioluminescence imaging of tumor, immunohistochemistry, flow cytometry, and cytokine multiplex assays to evaluate OMX’s ability to immunosensitize the GL261 brain tumor microenvironment and promote tumor cures. RESULTS: Following intravenous administration in brain tumor-bearing mice, OMX reduces tumor hypoxia, modulates the IFNg signaling pathway, enhances the infiltration of tumor-specific CX3CR1(+) CD8 T cells into the tumor (using the EphA2 as a GL261-specific tumor antigen), increases the activation of cytotoxic T lymphocytes (CTLs), decreases Tim3 and Lag3 exhaustion markers on CD8 T cells, and reduces the number of immunosuppressive cells such as MDSCs and Tregs in the tumor. Similar immunological changes are observed when OMX is combined with anti-PD-1. In late-stage tumor-bearing mice, we observed a 40% tumor cure rate for the combination of OMX with anti-PD-1, while anti-PD-1 alone resulted only in 5% tumor cures. Following rechallenge with GL261 tumor cells injected on the other side of the brain, all mice treated with the combination of OMX with anti-PD-1 survived, indicating the presence of long-term immunological memory against glioma cells. CONCLUSION: By delivering oxygen specifically to the hypoxic tumor microenvironment, OMX may restore anti-cancer immune responses in GB patients and synergize with radiotherapy and immunotherapy to enhance tumor control and improve patient outcomes.

There has been a change in the use of the au pair scheme in the past fifteen years that has created a shift from its original intention as a cultural exchange program. Socio- economic change in societies in the South and East has led to a new wave of female migrants seeking legal work opportunities in European countries, and change in the North has led to an increase in demand for domestic workers. The au pair program has become a means to cover these needs. Yet the use of the au pair institution as a temporary domestic work system creates challenges that not only contradict its intention, but also fail to offer labor rights and protection to the migrant women who enter the program to earn money. This article examines the au pair system in Norway, a country known for social and gender equality and a strongly developed welfare system based on social democratic ideals of solidarity. The article focuses in particular on how the au pair scheme is being misused as a temporary domestic work system by both the host families and the au pairs, and the exploitation and human trafficking cases that have emerged as a consequence in recent years.

Background: Hypoxia, a common feature in solid tumors such as glioblastoma (GB), is associated with resistance to chemo- and radio-therapies and poor patient outcomes. In addition, hypoxia promotes the immune escape of tumors. Therefore, reversing tumor hypoxia to create an immunopermissive microenvironment can improve antitumor response, and combined with immunotherapy approaches such as checkpoint inhibitors (CPI), may increase therapeutic efficacy. OMX is an oxygen carrier well tolerated in small (rats and mice) and large (sheep and dogs) animals. Following intravenous administration, OMX extravasates through leaky tumor vasculature and efficiently accumulates in orthotopic rodent GB and spontaneous canine brain tumors. Consequently, OMX significantly reduces hypoxia and improves the efficacy of radiotherapy and CPI. Methods: We used in vivo bioluminescence imaging of tumor, immunohistochemistry, flow cytometry, and cytokine multiplex assays to evaluate OMX's ability to immunosensitize the GL261 brain tumor microenvironment and promote tumor cures. Results: A single dose of OMX in brain tumor-bearing mice reduces tumor hypoxia, enhances the recruitment and infiltration of tumor-specific CX3CR1+ CD8 T cells into the tumor (using the EphA2 as a GL261-specific tumor antigen), decreases Tregs and increases activation and proliferation of cytotoxic T lymphocytes (CTLs). Specifically, OMX increases the Teff/Treg ratio by ~3-fold, indicating a switch from an immunosuppressive to an immunopermissive microenvironment. Similarly, when combined with anti-PD-1, OMX decreases Tregs, increases CTL infiltration, proliferation and cytotoxic activity, and modulates IFNg and IFNg-inducible cytokines that polarize T cells towards a Th1 phenotype. Treatment with OMX alone resulted in a 55% tumor cure rate, comparable to anti-PD-1 treatment. Furthermore, in late-stage tumor-bearing mice, we observed a 40% tumor cure rate for the combination of OMX with anti-PD-1, while anti-PD-1 alone resulted only in 5% tumor cures. In symptomatic mice with very high tumor burden, in which the combination of anti-PD-1 with anti-CTLA4 does not provide tumor cures, the addition of OMX resulted in a 20% tumor cure rate. Following rechallenge with GL261 tumor cells injected on the other side of the brain, all mice treated with OMX alone or in combination with CPI survived, indicating the presence of long-term immunologic memory against glioma cells. The survival benefit observed with OMX could be predicted with an identified circulating chemokine biomarker signature (post-hoc test). Conclusion: By delivering oxygen specifically to the hypoxic tumor microenvironment, OMX may restore anticancer immune responses in GB patients and synergize with radiotherapy and immunotherapy to enhance tumor control and improve patient outcomes. Citation Format: Natacha Le Moan, Philberta Leung, Sarah Ng, Tina Davis, Carol Liang, Jonathan Winger, Stephen P. Cary, Nicholas Butowski, Ana Krtolica. The oxygen carrier omx restores antitumor immunity and cures tumors as a single agent or in combination with checkpoint inhibitors in an intracranial glioblastoma mouse model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4726A.

Introduction: After vascular occlusion, there are two major zones of injury: the infarct core that rapidly dies, and the surrounding penumbra, which is hypoxic and at risk for infarction. Restoring blood flow by vascular recanalization (ie. endovascular +/- IV tPA) to prevent the penumbra from infarcting has become the new standard of care for acute ischemic stroke patients presenting with salvageable tissue. However, not all patients benefit from recanalization. Regardless of successful or partial reperfusion, secondary hypoxic events within the rescued penumbra can cause selective neuronal loss (SNL) that may account for suboptimal clinical recovery. OMX is a breakthrough oxygen delivery protein tuned to release oxygen specifically in hypoxic tissue and reduce hypoxia to prevent SNL within the reperfused penumbra. Methods: To mimic the clinical situation whereby occlusion is followed by gradual reperfusion after spontaneous or therapeutic thrombolysis, we used the endothelin-1 induced middle cerebral artery occlusion (MCAO) model. OMX was administered intravenously up to 2h post-MCAO. Short and long-term histological and behavioral outcomes were used to assess hypoxia, apoptosis, gliosis, SNL, infarct volume and sensorimotor functions. Results: Despite reperfusion, hypoxic tissue persists and progressively infarcts in vehicle-treated rats. However, OMX-treated rats showed no infarct expansion over 7d, indicating that OMX prevents infarction of the majority of hypoxic tissue. OMX reduces hypoxia and caspase 3/9 activity in a dose-dependent manner and significantly prevents SNL and gliosis. When administered post-MCAO, OMX significantly improves both sensory and motor functions (∼30-80% improvement, * p Conclusions: OMX is a promising therapeutic candidate that can be administered in combination with reperfusion therapies to stabilize the at-risk hypoxic tissue and ameliorate long-term clinical outcomes in stroke patients.

Oxygen is one of the key modulators of tumor microenvironment whereby low oxygen or hypoxia is associated with resistance to chemo- and radio- therapies and poor patient outcomes. Hypoxia favors an immunosuppressive tumor microenvironment by promoting Treg recruitment and activation and suppressing T cell and NK cell proliferation and effector function and pro-inflammatory cytokine secretion. Therefore, reversing tumor hypoxia could create an immunopermissive microenvironment and improve the efficacy of several immunotherapies. Omniox has developed an oxygen carrier OMX that can specifically deliver oxygen to hypoxic tumor regions without affecting oxygenation of tissues within physiologic oxygen levels. Due to its biochemical features, OMX is well tolerated in small (rats and mice) and large (sheep and dogs) animals. Following intravenous administration, OMX extravasates through leaky tumor vasculature and accumulates within immunocompetent rodent orthotopic glioblastoma models as well as spontaneous canine brain tumors. Consequently, OMX decreases hypoxia levels in the tumor tissue measured directly using oxygen sensor probes and indirectly with exogenous hypoxia markers using ELISA, immunohistochemistry and flow cytometry methods. Here we evaluated OMX’ activity in reversing the immunosupressive tumor microenvironment using a combination of immunohistochemistry, flow cytometry and Luminex methods. Moreover, we investigated the efficacy of OMX in improving mouse survival and effectiveness of checkpoint inhibitors (CPI). Similar to previously published findings, we demonstrated that T lymphocytes are mostly excluded from hypoxic tumor areas in the GL261 model. A single OMX treatment in GL261 tumor-bearing mice reduces tumor hypoxia, enhances T cell localization in previously hypoxic tumor areas, and increases CD8 accumulation by ~4-fold. Specifically, OMX treatment increased the activated cytotoxic T lymphocytes (CTLs) fraction by ~2 fold and reduced the immunosuppressive Treg fraction by 2-fold, resulting in a 3-fold increase of Teff/Treg ratio, which indicates a switch from an immunosupressive to an immunopermissive microenvironment. When combined with CPI, OMX reverses the immunosuppressive tumor microenvironment by increasing CD8 T cell infiltration, proliferation and cytotoxic activity, and modulating IFNg and IFNg-inducible cytokines that may polarize T cells towards a Th1 phenotype. Furthermore, treatment of late-stage GL261 tumor-bearing mice with the combination of OMX-CPI increases mouse survival by 80%. By delivering oxygen specifically to the hypoxic tumor microenvironment, OMX may restore anti-cancer immune responses in glioblastoma patients and synergize with radiotherapy and immunotherapy to enhance tumor control and improve patient outcomes. Citation Format: Natacha Le Moan, Philberta Leung, Sarah Ng, Tina Davis, Carol Liang, Jonathan W. Winger, Stephen P. Cary, Nicolas Butowski, Ana Krtolica. Omx a hypoxia modulator reverses the immunosuppressive glioblastoma microenvironment by stimulating T cell infiltration and activation that results in increased number of long-term survivors [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 4686. doi:10.1158/1538-7445.AM2017-4686

BACKGROUND: Hypoxic cells in solid tumors are resistant to radiotherapy. Omniox has developed a novel class of oxygen carrier proteins termed H-NOX, engineered to deliver oxygen selectively into hypoxic tissues. Previous studies have shown that one of our H-NOX candidates, OMX-4.80, penetrates intracranial glioblastoma (GB) tumors in mice, and in a dose dependent manner decreases tumor hypoxia and extends survival when combined with radiotherapy. However, we discovered that the circulation half-life of OMX-4.80 is ∼1h in dogs, which may limit clinical utility. To improve the circulation half-life, we developed a PEGylated version of OMX-4.80 (OMX-4.80P). The aim of this study was to characterize the pharmacokinetic and safety profile of OMX-4.80P and examine its effect on tumor hypoxia. METHODS: After intravenous administration of OMX-4.80P to mice and rats bearing GB tumors, we collected tumors, blood and plasma, and analyzed OMX-4.80P pharmacokinetics, immunogenicity and its effects on blood cell counts and chemistries. We evaluated OMX-4.80P bio-distribution by immunohistochemistry and its effect on tumor hypoxia by ELISA of endogenous (HIF-1a) and exogenous (pimonidazole) hypoxia markers. RESULTS: Compared to OMX-4.80, OMX-4.80P has an increased stability and circulation half-life (30h vs. 1h), significantly longer tumor accumulation (48h vs. 4h), and is not immunogenic. Furthermore, OMX-4.80P crosses the blood tumor barrier in several different intracranial rodent GB models and reduces HIF-1a and pimonidazole accumulation by ∼50%. Pharmacokinetic and toxicology studies in rodents and dogs showed that OMX-4.80P is well tolerated, and has a circulation half-life of ∼52h in dogs. The resulting exposure is expected to be sufficient for OMX-4.80P to accumulate in human GB tumors and reduce hypoxia. CONCLUSIONS: In summary, the increased retention of OMX-4.80P in circulation, the longer exposure achieved in solid tumors and efficiency in delivering oxygen into hypoxic tissues suggest OMX-4.80P is a promising IND candidate for radiation therapy enhancement in GB.

BACKGROUND: Hypoxia in solid tumors blunts the tumor cytotoxicity of radiotherapy (RT) and is a major independent predictor of poor patient outcomes in glioblastoma (GB). To increase tumor oxygenation, Omniox has engineered a novel oxygen delivery protein, OMX-4.80P (OMX), tuned to release oxygen specifically in hypoxic tumor tissue. When given with radiotherapy, OMX significantly enhances RT efficacy in rodent tumor models. Since spontaneous canine brain cancers, including GB, share similarities with human brain cancers in terms of morphology, disease progression, and response to approved therapies, a Phase 0 clinical trial was conducted in patient dogs to assess OMX safety, PK/PD and signs of efficacy to inform human clinical strategy. METHODS: Dogs with presumed brain cancers (MRI) underwent either resection surgery with radiotherapy (Sx + RT) or stereotactic radiosurgery (SRS). In the Sx + RT group, OMX was administered once before surgery (4–48 hours pre-Sx) and twice weekly in conjunction with RT (20x2.5Gy). In the SRS group, OMX was dosed on 2 consecutive days in combination with SRS (3x8Gy). PK, CBC, blood chemistry, coagulation, and immune responses were assessed. Additionally, resected tumors were analyzed for OMX accumulation and hypoxia levels (ELISA, IHC). RESULTS: To date, 14 dogs have completed the trial (10 Sx, 5 Sx + RT, 4 SRS): Five dogs show no signs of tumor regrowth at 3 months and 2 dogs had no visible tumors at 6 months. OMX induced no dose-limiting adverse events and median T1/2 is 35h. Resected tumors show a strong correlation between OMX concentration in tumor tissue and reduced hypoxia with OMX penetrating deep into tumor tissue. CONCLUSIONS: When given in conjunction with RT regimens equivalent to human standard of care, OMX is safe and well-tolerated. Tumor tissue analysis suggests that OMX-4.80P decreases tumor hypoxia, thus supporting its further clinical development for the enhancement of radiotherapy efficacy in GB patients.

BACKGROUND: Hypoxia is a hallmark of glioblastoma (GB) and is correlated with tumor progression and decreased overall survival. OMX-4.80P is a protein oxygen carrier engineered for the treatment of GB in combination with radiotherapy. Prior studies in rodent GB models demonstrated that OMX-4.80P accumulates within intracranial tumors and reduces tumor hypoxia—as assessed by immunohistochemical and ELISA methods—and enhances tumor killing by radiotherapy. To support clinical evaluation of OMX-4.80P in patients, this report describes non-invasive positron emission tomography (PET) imaging to assess both OMX-4.80P tumor accumulation and oxygen delivery. METHODS: To assess its biodistribution, OMX-4.80P was labelled with 89Zr using a desferrioxamine (DFO) chelator. Either 89Zr-DFO-OMX-4.80P or free 89Zr was administered intravenously to both nude mice bearing H460 subcutaneous xenografts, as well as to Fischer rats bearing intracranial F98 tumors. OMX-4.80P distribution was assessed by PET, and by measuring radioactivity in major organs using a gamma counter. To assess OMX-4.80P-mediated changes in tumor hypoxia in the same tumor models, tumor:brain signal ratio of the hypoxia marker fluoromisonidazole (18F-FMISO) was assessed before and after treatment with OMX-4.80P by performing scans on two consecutive days. RESULTS: While free 89Zr was present mainly in the bones, 89Zr-DFO-OMX-4.80P was not, suggesting that the label is stable in vivo. As expected, 89Zr-DFO-OMX-4.80P accumulated in both subcutaneous and intracranial tumors, and in the clearance organs (liver, kidneys, spleen). Furthermore, 18F-FMISO levels, as measured by tumor:brain ratios, were decreased by >2 fold after either intravenous or intratumoral injection of OMX-4.80P, in both subcutaneous and intracranial tumors demonstrating OMX-4.80P-mediated tumor oxygenation activity. CONCLUSIONS: PET-based approaches are a feasible method for assessing OMX-4.80P distribution and oxygenation. These methods will be developed as biomarkers of OMX-4.80P activity in a planned Phase 1 clinical trial in newly diagnosed GB patients.

Familial hypercholesterolemia (FH) remains a significantly undertreated genetic disease in the United States despite the availability of effective therapies (1). Without intervention, patients face high morbidity and mortality rates; however, early diagnosis and treatment to guidelines can reduce event rates to those found in the general population (2). Each FH1 and FH2 patient was tagged in the EHR to increase visibility to all providers. Providers received system-wide communiques about the program. Total number of patients identified is outlined in Figure A. Synopsis

BACKGROUND: Omniox has engineered OMX-4.80P, a PEGylated H-NOX oxygen carrier, as a long-acting therapeutic candidate to enhance radiotherapy (RT) in the treatment of glioblastoma (GB) and other solid tumors. We have previously described the un-PEGylated OMX-4.80 variant, which can penetrate intracranial mouse GB tumors, release oxygen exclusively into hypoxic tumor tissue, and enhance tumor growth delay and survival when combined with RT. However, OMX-4.80 has a short circulation half-life that limits its clinical potential. Here, we describe the pre-clinical profile of PEGylated OMX-4.80 (OMX-4.80P), demonstrating it is well tolerated, long-lasting in circulation and tumors, and is significantly more efficacious than OMX-4.80. METHODS: We PEGylated OMX-4.80 to increase its circulation half-life while retaining its ability to preferentially accumulate in tumors and selectively release oxygen to hypoxic microenvironments. We characterized the efficacy of OMX-4.80P in large, hypoxic, radioresistant tumors, and its activity in intracranial GB models in nude mice and immunocompetent rats. We also conducted toxicology and pharmacokinetic studies in multiple animal models including naïve dogs. RESULTS: In xenograft studies of large, hypoxic, radioresistant tumors, single doses of OMX-4.80P in combination with RT result in apparent tumor cures in ∼30-50% of tumors compared to 0% cures in RT-only groups. Interestingly, the larger molecular size of OMX-4.80P (∼120kDa) did not prevent its passing the blood-tumor barrier in mouse and rat intracranial tumors and resulted in hypoxia reduction. Pharmacokinetic and toxicology studies using single or multiple supratherapeutic and therapeutic doses of OMX-4.80P in rodents and dogs demonstrated that it has a circulation half-life of ∼24h in rats and ∼52h in dogs, compared to 1-2h for OMX-4.80, and that it is well tolerated. Finally, OMX-4.80P has no detectable immunogenic response. CONCLUSIONS: The preclinical efficacy data, and promising pharmacokinetic and toxicology profile, of OMX-4.80P support its clinical development for the enhancement of radiotherapy in GB patients.

Background and Purpose —The purpose of this study was to assess the efficacy of neuromuscular stimulation in enhancing the upper extremity motor and functional recovery of acute stroke survivors. Methods —Forty-six stroke survivors admitted to an inpatient rehabilitation unit were randomly assigned to receive either neuromuscular stimulation or placebo. Twenty-eight subjects completed the study. The treatment group received surface neuromuscular stimulation to produce wrist and finger extension exercises. The control group received placebo stimulation over the paretic forearm. All subjects were treated 1 hour per day, for a total of 15 sessions. Outcomes were assessed in a blinded manner with the upper extremity component of the Fugl-Meyer Motor Assessment and the self-care component of the Functional Independence Measure at pretreatment, after treatment, and at 4 and 12 weeks after treatment. Results —The treatment subjects and control subjects had comparable baseline characteristics. Parametric analyses revealed significantly greater gains in Fugl-Meyer scores for the treatment group after treatment (13.1 versus 6.5; P =0.05), at 4 weeks after treatment (17.9 versus 9.7; P =0.05), and at 12 weeks after treatment (20.6 versus 11.2; P =0.06). Functional Independence Measure scores were not different between groups at any of the time periods ( P >0.10). Conclusions —Data suggest that neuromuscular stimulation enhances the upper extremity motor recovery of acute stroke survivors. However, the sample size in this study was too small to detect any significant effect of neuromuscular stimulation on self-care function.

Purpose Radiation dermatitis is a common side effect of radiation therapy (RT). In severe cases, RT must be interrupted until the skin heals, which can compromise treatment. The purpose of the study was to compare an anionic polar phospholipid (APP)-based cream and an aloe vera-based gel to determine their effectiveness in preventing and treating radiation dermatitis. Patients and methods Forty-five pediatric patients (median age, 11 years) with various diagnoses who received at least 23.4 Gy participated. APP cream and aloe vera gel were symmetrically applied within the irradiated field after each treatment. Three measures were collected before, during and after completion of treatment: subject's skin comfort, dermatologic assessment, and common toxicity criteria (CTC). Results Significant differences in specific variables favoring APP cream use were noted in some patients including skin comfort variables, dry (p = 0.002), soft (p = 0.057), feels good (p = 0.002), rough (p = 0.065), smooth (p = 0.012) and dermatologic variables, dryness (p = 0.013), erythema (p = 0.002) and peely (p = 0.008). Grouped CTC scores were supportive of APP cream (p = 0.004). In comparing the first and last assessments, two dermatologic variables, dryness (p = 0.035) and peely (p = 0.016), favored APP cream. Conclusion APP cream is more effective than aloe vera-based gel for prevention and treatment of radiation dermatitis.