Therapeutic Efficacy Of Combination Therapy Research Articles

Novel metallo-β-lactamases inhibitors restore the susceptibility of carbapenems to New Delhi metallo-lactamase-1 (NDM-1)-harbouring bacteria.

The production of metallo-β-lactamases is a major mechanisms adopted by bacterial pathogens to resist carbapenems. Repurposing approved drugs to restore the efficacy of carbapenems represents an efficient and cost-effective approach to fight infections caused by carbapenem resistant pathogens. The nitrocefin hydrolysis assay was employed to screen potential New Delhi metallo-lactamase-1 (NDM-1) inhibitors from a commercially available U.S. Food and Drug Administration (FDA) approved drug library. The mechanism of inhibition was clarified by metal restoration, inductively coupled plasma mass spectrometry (ICP-MS) and molecular dynamics simulation. The in vitro synergistic antibacterial effect of the identified inhibitors with meropenem was determined by the checkerboard minimum inhibitory concentration (MIC) assay, time-dependent killing assay and combined disc test. Three mouse infection models were used to further evaluate the in vivo therapeutic efficacy of combined therapy. Twelve FDA-approved compounds were initially screened to inhibit the ability of NDM-1 to hydrolyse nitrocefin. Among these compounds, dexrazoxane, embelin, candesartan cilexetil and nordihydroguaiaretic acid were demonstrated to inhibit all tested metallo-β-lactamases and showed an in vitro synergistic bactericidal effect with meropenem against metallo-β-lactamases-producing bacteria. Dexrazoxane, embelin and candesartan cilexetil are metal ion chelating agents, while the inhibition of NDM-1 by nordihydroguaiaretic acid involves its direct binding to the active region of NDM-1. Furthermore, these four drugs dramatically rescued the treatment efficacy of meropenem in three infection models. Our observations indicated that dexrazoxane, embelin, candesartan cilexetil and nordihydroguaiaretic acid are promising carbapenem adjuvants against metallo-β-lactamases-positive carbapenem resistant bacterial pathogens.

Enhanced Systemic Antitumour Immunity by Hypofractionated Radiotherapy and Anti-PD-L1 Therapy in Dogs with Pulmonary Metastatic Oral Malignant Melanoma.

Although immune checkpoint inhibitors (ICIs), such as the anti-programmed death-ligand 1 (PD-L1) antibody, have been developed for the treatment of canine malignant melanoma, desirable clinical efficacies have not been achieved. Recent studies in humans have suggested that radiation therapy (RT) combined with ICIs induces robust systemic antitumour immunity in patients with cancer. This study retrospectively examined the therapeutic efficacy of combination therapy (hypofractionated RT and anti-PD-L1 antibody [c4G12]) in dogs with pulmonary metastatic oral malignant melanoma. The intrathoracic clinical benefit rate (CBR)/median overall survival (OS) in the no RT (n = 20, free from the effect of RT), previous RT (n = 9, received RT ≤8 weeks prior to the first c4G12 dose), and concurrent RT (n = 10, c4G12 therapy within ±1 week of the first RT fraction) groups were 10%/185 days, 55.6%/283.5 days (p < 0.05 vs. no RT group), and 20%/129 days (p > 0.05 vs. no RT group), respectively. The adverse events were considered to be tolerable in the combination therapy. Thus, hypofractionated RT before the initiation of c4G12 therapy can be an effective approach for enhancing the therapeutic efficacy of immunotherapy, with acceptable safety profiles. Further prospective clinical studies are required to confirm the findings of this study.

Клинико-патогенетические аспекты лечения и профилактики пиелонефрита у детей

Treatment of pyelonephritis in children should be combined, long-term and individual-based. The success of the therapy in children largely depends on the prompt appointment and the correct choice of antimicrobial therapy.To evaluate the efficiency of the dietary supplement "Cystenium II" in a group of children aged 7 to 14 years with a diagnosis of acute and chronic recurrent pyelonephritis in the acute phase.A total of 60 children aged 7 to 14 years with a diagnosis of acute or chronic recurrent pyelonephritis in the acute stage were included in the study. The clinical group consisted of 30 patients (mean age 12.1+/-1.8 years), while the control group included 30 patients of mean age 11.2+/-1.7 years. In the control group patients received only standard antibiotic therapy, while in the clinical group it was combined with a dietary supplement "Cystenium II" 1 tablet 2 times a day with meals for 14 days. After the course of antibacterial treatment, the children in the clinical group continued to take the studied dietary supplement for another 14 days in order to prevent the recurrence of pyelonephritis. The results of treatment (patient's condition, presence of pain, dysuria, fever) were assessed on the 3rd, 7th, 14th day, 1 and 6 months after the start of treatment. A urinalysis was performed at the baseline, on the 7th and 14th days, as well as after 1 and 6 months. Urine culture was performed before and after antibiotic therapy at the baseline, on the 14th day, 1 and 6 months after the start of treatment.The main indicators of urinalysis (leukocytes, red blood cells, protein) returned to normal values in 26 (86.7%) patients of the clinical group and in 23 (76.7%) patients of the control group on the 7th day after the start of treatment. At the completion of the basic therapy (after 14 days) normal clinical parameters (absence of leukocyturia, microhematuria, proteinuria) were observed in all patients of the clinical group and in 28 (93.3%) patients of the control group. After a month of follow-up, the disturbances in urinalysis (leukocytes, red blood cells, protein) in the control group were again seen in 3 (10%) patients, as well as after 6 months. However, in the clinical group all patients had normal urinalysis (absence of leukocyturia, microhematuria, proteinuria) after 1 month and only in 1 (3.3%) case leukocyturia, as well as an increase in the number of red blood cells and protein was detected by 6 months.According to our results, the use of dietary supplements "Cystenium II" (manufactured by Akvion, Russia), due to the constituents of D-mannose (450 mg), cranberry fruit extract with a standardized activity of 500 mg (36 mg of proanthocyanidins) and vitamin C (60 mg), may cause anti-inflammatory and anti-adhesive effects (resolving of leukocyturia and bacteriuria). This allows to use the dietary supplement Cystenium II in children from 7 years of age in the combination therapy of acute pyelonephritis, as well as exacerbation of chronic pyelonephritis. The obtained results showed a high overall therapeutic efficacy of combination therapy using Cystenium II after 6 months from the start of treatment (relapse in 1 patient), in contrast to the control group (relapse in 6 patients).the use of dietary supplement "Cystenium II" allowed to reduce the number of repeated courses of antibiotic therapy in children during 6 months of follow-up and, most likely, reduced the frequency of development of chronic pyelonephritis after an acute inflammation. Therefore, the wide clinical use of dietary supplements "Cystenium II" for the combined treatment of acute and exacerbation of chronic pyelonephritis in children older than 7 years seems to be very reasonable.

Pharmacokinetic and gut microbiota analyses revealed the effect of Lactobacillus acidophilus on the metabolism of Olsalazine in ulcerative colitis rats

Olsalazine is a typical 5-aminosalicylic acid (5-ASA) drug that depends on gut microbiota to liberate its anti-inflammatory moiety 5-ASA in the treatment of ulcerative colitis (UC). In recent decades, 5-ASA drugs combined with probiotics have achieved a better effective treatment for UC. Mechanisms of combination therapy have been widely discussed from a pharmacodynamic perspective. However, it is still unclear whether the better therapeutic efficacy of combination therapy was made by changing the metabolism of 5-ASA drugs in the colon under the regulation of probiotics. In the present study, combined with pharmacokinetic and gut microbiota analyses, we systematically evaluated the potential effect of Lactobacillus acidophilus (L. acidophilus) on the metabolism of Olsalazine at three levels (pharmacokinetic characteristics, metabolic microbiota, and metabolic enzymes) to offer some insights into this issue. As pharmacokinetic results showed, L. acidophilus barely had an influence on the pharmacokinetic parameters of Olsalazine, 5-ASA, and N-Ac-5-ASA. Notably, the colonic exposure of 5-ASA was not affected by L. acidophilus. Gut microbiota results also illustrated that L. acidophilus did not change the total abundance of azoreductase (azoR) and N-acetyltransferase (NAT) associated gut microbiota and enzymes, which are involved in the metabolism of Olsalazine. Both pharmacokinetic and gut microbiota results revealed that L. acidophilus did not increase the colonic exposure of 5-ASA to improve the efficacy of combination therapy. L. acidophilus played its role in UC treatment by regulating gut microbiota composition and amino acid, phenolic acid, oligosaccharide, and peptidoglycan metabolic pathways. There was no potential medication risk of combination therapy of Olsalazine and L. acidophilus. In summary, this research provided strong evidence of medication safety and a comprehensive understanding of therapeutic advantages for combination therapy of probiotics and 5-ASA drugs from the pharmacokinetic and gut microbiota perspectives.

Pervasive developmental disorders with a complex structure (epileptic seizures, catatonic, hallucinatory and manic symptoms, delirious episodes during life)

IntroductionThe significant clinical efficacy of lithium and lamotrigine in the described patients is consistent with the hypothesis that microdeletion of the SHANK3 gene may be associated with bipolar disorder.ObjectivesThe described phenotypes were characterized by intellectual disability, general speech underdevelopment, muscle hypotonia, and developmental dyspraxia. Their causal relationships with epileptic encephalopathy, schizophrenia, bipolar and hyperkinetic disorders have been analyzed.MethodsProspective studies of two patients with Phelan McDermid syndrome (22q13.3 microdeletions of the SHANK3 gene) within 10-12 years after genome scanning allowed describing the clinical polymorphism of developmental disorders. The sensitivity of psychotic symptoms to antipsychotic and mood stabilizer therapy has also been studied.ResultsManifest psychotic disorders in patients did not reveal an affinity for therapy with amisulpride, haloperidol, quetiapine, which demonstrated a partial therapeutic response to treatment with aripiprazole, which cast doubt on the possibility of qualifying psychotic symptoms in patients as the debut of schizophrenia. The partial therapeutic efficacy of combination therapy with aripiprazole and benzodiazepines (clonazepam/diazepam) qualified psychotic episodes in two patients with 22q13.3 syndrome as pediatric delirium. The significant clinical efficacy of lithium and lamotrigine in the described patients is consistent with the hypothesis that microdeletion of the SHANK3 gene may be associated with bipolar disorder.ConclusionsThe combination of lithium and lamotrigine may be recommended for the treatment of polymorbid mental disorders in patients with ASD and 22q13.3 syndrome. If lithium salts are poorly tolerated, a combination of lamotrigine and aripiprazole may be used to treat polymorbid mental disorders with confusion and catatonic symptoms in ASD.DisclosureNo significant relationships.

Exosomes derived from γδ-T cells synergize with radiotherapy and preserve antitumor activities against nasopharyngeal carcinoma in immunosuppressive microenvironment

BackgroundRadiotherapy is the first-line treatment for patients nasopharyngeal carcinoma (NPC), but its therapeutic efficacy is poor in some patients due to radioresistance. Adoptive T cell-based immunotherapy has also shown promise...

Pervasive developmental disorders with a complex structure (epileptic seizures, catatonic, hallucinatory and manic symptoms, delirious episodes during life).

Observation of two patients with Phelan-McDermid syndrome (22q13.3 microdeletions of the SHANK3 gene) within 10—12 years allowed us to describe the clinical pathomorphism of psychotic episodes with violation of consciousness and catatonic symptoms in adolescence with autism spectrum disorders (ASD). The described phenotypes were characterized by intellectual disability, general speech underdevelopment, muscle hypotonia and developmental dyspraxia. Their causal relationships with epileptic encephalopathy, schizophrenia, bipolar and hyperkinetic disorders have been analyzed. The therapeutic efficacy of combination therapy with aripiprazole and benzodiazepines (clonazepam/diazepam) allowed qualifying psychotic episodes as pediatric delirium. The significant clinical efficacy of lithium and lamotrigine in the described patients was consistent with the hypothesis that microdeletion of the SHANK3 gene may be associated with bipolar disorder. Treatment of acute psychotic disorders with lithium salt was effective in both patients but had limitations due to poor tolerance in the long-term use. The combination of lithium and lamotrigine may be recommended for the treatment of polymorbid mental disorders in patients with SHANK3 encephalopathies. If lithium salts are poorly tolerated, a combination of lamotrigine and aripiprazole may be used.

1% ivermectin in combination therapy regimens for rosacea

Introduction. The article provides latest data on modern methods of treating rosacea. The results of own clinical observations of patients with moderate to severe papulopustular rosacea receiving combination treatment and a comparative analysis of the efficacy of various therapy regimens are presented.Objective of the study. The aim of the study was to conduct a comparative analysis of the therapeutic efficacy of combination therapy using the ivermectin 1% topical drug combined with systemic therapy drugs (doxycycline, minocycline, isotretinoin).Materials and methods. We observed 37 patients with moderate to severe papulo-pustular rosacea subtype. The patients were divided into 4 groups (A, B, C, D). Patients in the control group received monotherapy with 1% ivemectin topical drug, patients in the other groups received combination therapy (1% ivermectin combined with low-dose doxycycline, minocycline and isotretinoin). The efficacy of the therapy was evaluated by measuring rosacea severity on the Scale for Diagnostic Assessment of Rosacea (SDAR), clinical manifestations according to the IGA (Investigator Global Assessment) criteria, and by assessing the patients' quality of life using the DLQI (Dermatology Life Quality Index) questionnaire before and after 3-month treatment.Results. The comparative analysis of changes in severity indicators of the skin process and quality of life in patients with moderate to severe papulopustular rosacea after topical and combination therapy showed that the results of the treatment in patients receiving combination therapy were more significant than those in the group receiving monotherapy.Conclusion. The concomitant use of 1% ivermectin and systemic drugs is most effective in patients with severe papulopustular rosacea subtype. The combination treatment tailored to the clinical forms and severity of rosacea allows to optimize the clinical results of the therapy, which significantly affects the patients' quality of life and opens up potential for an individual approach in the algorithms for the treatment of rosacea.

Abstract 562: Novel combination therapy for treating TNBC using LIFR and HDAC Inhibitors

Abstract Background: Triple-negative breast cancer (TNBC) lacks targeted therapies and represents a disproportional share of the breast cancer (BC) mortality rate. TNBC exhibits autocrine stimulation of the LIF/LIFR axis and overexpression of LIF is associated with poorer relapse-free survival in BC patients. Histone deacetylase inhibitors (HDACIs) are emerging as promising multifunctional agents in TNBC to elicit cytotoxic actions. Recent studies have shown that cancer cells elicit feedback activation of leukemia inhibitory factor receptor (LIFR) which in turn curtails response to HDACIs. We developed a first-in-class inhibitor of LIFR, EC359 that directly interacts with LIFR and effectively blocks LIFR downstream signaling. The objective of this study is to examine the therapeutic efficacy of combination therapy using preclinical and patient-derived xenograft (PDX) models. Methods: We tested utility of combination therapy using multiple HDACIs that are currently in clinical trails along with EC359. The effect of combination therapy was evaluated using MTT, invasion, colony formation, and Caspase3/7 assays. Mechanistic studies were performed using Western blotting, qRT-PCR, and STAT3 reporter assays. The efficacy of combination therapy in vivo was examined using xenograft, PDX, and patient-derived explant (PDEx) models. Results: Immunohistochemical analyses of breast tumors using tissue microarrays revealed significant expression of LIFR in TNBC tissues. Treatment of TNBC model cells with four different HDACIs increased the expression of LIFR. LIFR inhibitor EC359 at nM concentration is additive to HDACIs in reducing cell viability. Knockdown of LIFR or treatment with EC359 significantly enhanced the efficacy of HDACIs in reducing the cell viability, colony formation ability, and invasiveness as well as promoted apoptosis compared to monotherapy in TNBC model cells. On the contrary, treatment with STAT3 inhibitor requires µM concentrations to reduce the cell viability of TNBC cells and is not additive to HDACIs. Mechanistic studies utilizing STAT3 reporter gene assays and biochemical studies using multiple TNBC model cells exhibited activation of the LIFR signaling pathway upon HDACIs treatment but was attenuated by EC359 therapy. Treatment of human TNBC utilizing PDEx assays showed that EC359 enhanced the ability of HDACIs to decrease proliferation (Ki-67 positivity) compared to monotherapy. Using TNBC xenografts and PDX models, we demonstrated that EC359 treatment enhanced the ability of HDACIs to reduce in vivo tumor growth compared to monotherapy. Conclusions: Our results suggest that the combination therapy of HDACIs and EC359 provides therapeutic utility in overcoming the limitation of feedback activation of LIFR observed in the treatment of HDACIs in treating TNBC. Supported by DOD BCRP grant W81XWH-18-1-0016 (R.K. Vadlamudi; K.J. Nickisch) Citation Format: Suryavathi Viswanadhapalli, Mengxing Li, Bindu Santhamma, Uday P. Pratap, Yiliao Luo, Junhao Liu, Kristin A. Altwegg, Xiaonan Li, Ahmed Gulzar, Hui Yan, Zhenming Xu, Andrew Brenner, Gangadhara R. Sareddy, Manjeet K. Rao, Rajeshwar R. Tekmal, Hareesh B. Nair, Klaus J. Nickisch, Ratna K. Vadlamudi. Novel combination therapy for treating TNBC using LIFR and HDAC Inhibitors [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 562.

Abstract 3732: Targeted photodynamic therapy enhances the therapeutic efficacy of combination therapy (PDT and chemotherapy) on chemoresistant melanoma cells

Abstract Cutaneous melanoma remains refractory to current therapies such as chemotherapy. Failure of treatment has been attributed to enhanced self-renewal capacity and the over-expression of ABC transporters by therapy resistant populations. A need therefore exists to identify and evaluate new approaches. Alternative combinatorial approaches comprising chemotherapeutics, photodynamic therapy (PDT) and targeted-PDT have been proposed to overcome resistance. Objective This study set to investigate the efficacy of novel (targeted-PDT-IR700) and standard combinatorial effect of PDT and chemotherapy (Hypericin-PDT and DTIC) as an adjunctive combination therapy treatment to decrease cellular chemoresistance in melanoma. This was assessed in an in vitro model, to show differences in killing of melanoma cells. Methodology Single chain variable fragments (scFv) of panitumumab were cloned into a transient eukaryotic SNAP expression vector co-expressing cytosolic proteins and transfected into HEK293T cells. Purified recombinant protein was confirmed by SDS-PAGE and Western blot analyses. Benzylguanine (BG) modified IR700 was irreversibly conjugated in a 1:1 stochiometric ratio to the 1711(scFv)-SNAP using the self-labelling property of the suicide enzyme SNAP-tag. Binding activity and cytotoxicity was documented on EGFR expressing melanoma cells using flow cytometry, confocal microscopy and XTT assays. Cytotoxic activity, self-renewal capacity and ABC transporters expression (ABCB5 and ABCG2) after the combination therapy (HYP-PDT and DTIC) was determined using XTT, clonogenic assays and flow cytometry. Results Binding and internalization of purified 1711(scFv)-SNAP conjugated to BG-modified Alexa 488 or IR700 was successfully confirmed by confocal microscopy and flow cytometry. Cell viability showed cell specific cytotoxicity of the photosensitizer labeled fusion proteins upon light induction, at nanomolar concentrations which were lower than those used for the non-targeted combinatorial approach (HYP-PDT and DTIC). Moreover, DTIC resistant melanoma cells were less sensitive to therapeutic treatment than their non-resistant counterpart. This correlated with an increased self-renewal capacity and ABC transporter expression in DTIC resistant melanoma in comparison to their non-resistant counterpart. This study describes a proof of concept of selective targeting and elimination of EGFR expressing melanoma cells by targeted photoimmunotherapy using a panitumumab 1711-scFv fused to SNAP tag and conjugated to near infrared photosensitizer IR700 which improve therapeutic efficacy with lower concentration and reduced sides effect. This work highlights the efficacy of targeted PDT and suggest that it is highly applicable to improve the current clinical arsenal of therapies. Note: This abstract was not presented at the meeting. Citation Format: Lester M. Davids, Fleury Nsole Biteghe, Eden Padayachee, Stefan Barth. Targeted photodynamic therapy enhances the therapeutic efficacy of combination therapy (PDT and chemotherapy) on chemoresistant melanoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3732.

A Mixture of U.S. Food and Drug Administration-Approved Monoaminergic Drugs Protects the Retina From Light Damage in Diverse Models of Night Blindness.

PurposeThe purpose of this study was to test the extent of light damage in different models of night blindness and apply these paradigms in testing the therapeutic efficacy of combination therapy by drugs acting on the Gi, Gs, and Gq protein-coupled receptors.MethodsAcute bright light exposure was used to test susceptibility to light damage in mice lacking the following crucial phototransduction proteins: rod transducin (GNAT1), cone transducin (GNAT2), visual arrestin 1 (ARR1), and rhodopsin kinase 1 (GRK1). Mice were intraperitoneally injected with either vehicle or drug combination consisting of metoprolol (β1-receptor antagonist), bromocriptine (dopamine family-2 receptor agonist) and tamsulosin (α1-receptor antagonist) before bright light exposure. Light damage was primarily assessed with optical coherence tomography and inspection of cone population in retinal whole mounts. Retinal inflammation was assessed in a subset of experiments using autofluorescence imaging by scanning laser ophthalmoscopy and by postmortem inspection of microglia and astrocyte activity.ResultsThe Gnat1−/− mice showed slightly increased susceptibility to rod light damage, whereas the Gnat2−/− mice were very resistant. The Arr1−/− and Grk1−/− mice were sensitive for both rod and cone light damage and showed robust retinal inflammation 7 days after bright light exposure. Pretreatment with metoprolol + bromocriptine + tamsulosin rescued the retina in all genetic backgrounds, starting at doses of 0.2 mg/kg metoprolol, 0.02 mg/kg bromocriptine, and 0.01 mg/kg tamsulosin in the Gnat1−/− mice. The therapeutic drug doses increased in parallel with light-damage severity.ConclusionsOur results suggest that congenital stationary night blindness and Oguchi disease patients can be at an elevated risk of the toxic effects of bright light. Furthermore, systems pharmacology drug regimens that stimulate Gi signaling and attenuate Gs and Gq signaling present a promising disease-modifying therapy for photoreceptor degenerative diseases.

Astragalus membranaceus improves therapeutic efficacy of asthmatic children by regulating the balance of Treg/Th17 cells

Astragalus membranaceus may be a potential therapy for childhood asthma but its driving mechanism remains elusive. The main components of A. membranaceus were identified by HPLC. The children with asthma remission were divided into two combination group (control group, the combination of budesonide and terbutaline) and A. membranaceus group (treatment group, the combination of budesonide, terbutaline and A. membranaceus). The therapeutic results were compared between two groups after 3-month therapy. Porcine peripheral blood mononuclear cells (PBMCs) were isolated from venous blood by using density gradient centrifugation on percoll. The levels of FoxP3, EGF-β, IL-17 and IL-23 from PBMCs and serum IgE were measured. The relative percentage of Treg/Th17 cells was determined using flow cytometry. The main components of A. membranaceus were calycosin-7-O-glucoside, isoquercitrin, ononin, calycosin, quercetin, genistein, kaempferol, isorhamnetin and formononetin, all of which may contribute to asthma therapy. Lung function was significantly improved in the treatment group when compared with a control group (P < 0.05). The efficacy in preventing the occurrence of childhood asthma was higher in the treatment group than the control group (P < 0.05). The levels of IgE, IL-17 and IL-23 were reduced significantly in the treatment group when compared with the control group, while the levels of FoxP3 and TGF-β were increased in the treatment group when compared with the control group (P < 0.05). A. membranaceus increased the percentage of Treg cells and reduced the percentage of Th17 cells. A. membranaceus is potential natural product for improving the therapeutic efficacy of combination therapy of budesonide and terbutaline for the children with asthma remission by modulating the balance of Treg/Th17 cells.

Therapeutic Efficacy of Combination Therapy Using Oral Cyclosporine with a Dietary Supplement (Pantogar®) in Twenty-Nail Dystrophy.

BackgroundTwenty-nail dystrophy (TND) is an acquired idiopathic disease characterized by dull, thin, lusterless, and fragile nails with fissuring, small regular pits, and excessive longitudinal ridging. Although various treatment modalities have been performed in order to treat TND, the effects of these treatments are controversial.ObjectiveTo evaluate the effectiveness of oral cyclosporine in TND.MethodsA total of 38 patients with TND were treated with combination therapy using oral cyclosporine with a pantothenic acid complex-based dietary supplement (Pantogar®; Merz Pharmaceuticals GmbH, Germany), whereas 44 patients were treated with the pantothenic acid complex-based dietary supplement alone. The therapeutic efficacy in each group was retrospectively evaluated using medical records and clinical photographs.ResultsThe cyclosporine therapy group had significantly more patients whose improvement was rated as almost clear, marked, or moderate compared to the control group (p<0.001). While the mean CHATS (Color, Hyperkeratosis, Area, Thickness, Separation) score of the cyclosporine therapy group was decreased by 13.45 (from 30.95 to 17.5) after treatment, the mean CHATS score of the control group was only decreased by 8 (from 29.43 to 21.43, p<0.001). Moreover, greater Dermatology Life Quality Index changes after treatment were observed in the cyclosporine therapy group (p=0.085).ConclusionOral cyclosporine can be a valuable therapeutic option in patients with TND.

Highlights of This Issue

Low socioeconomic status (SES) is associated with adverse outcomes among unrelated donor hematopoietic stem cell transplant (HCT) recipients, but the biological mechanisms contributing to this health disparity are poorly understood. Knight and colleagues found that low-SES HCT recipients show up-regulation of a conserved transcriptional response to adversity (CTRA) gene expression profile at pretransplant testing. Up-regulated CTRA gene expression was also associated with increased relapse and decreased leukemia-free survival. These results suggest new strategies for mitigating social disparities in cancer treatment outcomes through targeted pharmacologic intervention in the signaling pathways that drive CTRA gene expression.Hatzis and colleagues developed a statistical model to help design the next generation of clinical trials for breast cancer that simultaneously test drug activity in the preoperative (neoadjuvant) treatment setting and the impact on patient survival. Neoadjuvant chemotherapy often completely eradicates the cancer from the breast and lymph nodes, but does not always result in improved survival. The team found higher expected survival benefit for cancers with high risk of recurrence when treated with more effective novel therapies, highlighting the importance of assessing novel treatments in patients that have high risk for recurrence despite current best therapies.Combinations of ibrutinib with anti-CD20 monoclonal antibodies (mAbs) are being evaluated in clinical trials; however, in vitro data identified potentially negative interactions between the two drugs. Skarzynski and colleagues provide conclusive evidence that ibrutinib leads to CD20 downregulation on chronic lymphocytic leukemia (CLL) cells in vivo, resulting in decreased complement-dependent cytotoxicity. Yet, concurrent CD55 (complement inhibitor) downregulation and partial inhibition of mAb-mediated shaving of CD20 from CLL cells by effector cells (trogocytosis) could enhance mAb activity. These positive and negative interactions between ibrutinib and anti-CD20 mAbs could determine the overall therapeutic efficacy of combination therapy and should be investigated further.The site of recurrence affects prognosis in metastatic breast cancer, but the events underlying organ-specificity of metastases are elusive. The authors explored the global transcriptional landscape of clinical breast cancer metastases, unraveling a liver metastasis-selective signature enriched for stromal genes that were predominantly downregulated in liver metastases. Downregulation of the signature was predictive of poor outcome in early breast cancer, with the strongest independent prognostic value within the luminal A subtype. This signature identifies patients with ER-positive breast cancer requiring closer disease monitoring who may be amenable to enrolment into clinical trials investigating novel therapeutics targeting features other than proliferation.

Optimal treatment scheduling of ionizing radiation and sunitinib improves the antitumor activity and allows dose reduction.

The combination of radiotherapy with sunitinib is clinically hampered by rare but severe side effects and varying results with respect to clinical benefit. We studied different scheduling regimes and dose reduction in sunitinib and radiotherapy in preclinical tumor models to improve potential outcome of this combination treatment strategy. The chicken chorioallantoic membrane (CAM) was used as an angiogenesis in vivo model and as a xenograft model with human tumor cells (HT29 colorectal adenocarcinoma, OE19 esophageal adenocarcinoma). Treatment consisted of ionizing radiation (IR) and sunitinib as single therapy or in combination, using different dose-scheduling regimes. Sunitinib potentiated the inhibitory effect of IR (4 Gy) on angiogenesis. In addition, IR (4 Gy) and sunitinib (4 days of 32.5 mg/kg per day) inhibited tumor growth. Ionizing radiation induced tumor cell apoptosis and reduced proliferation, whereas sunitinib decreased tumor angiogenesis and reduced tumor cell proliferation. When IR was applied before sunitinib, this almost completely inhibited tumor growth, whereas concurrent IR was less effective and IR after sunitinib had no additional effect on tumor growth. Moreover, optimal scheduling allowed a 50% dose reduction in sunitinib while maintaining comparable antitumor effects. This study shows that the therapeutic efficacy of combination therapy improves when proper dose-scheduling is applied. More importantly, optimal treatment regimes permit dose reductions in the angiogenesis inhibitor, which will likely reduce the side effects of combination therapy in the clinical setting. Our study provides important leads to optimize combination treatment in the clinical setting.

Therapeutic efficacy of combination therapy with intra-arterial 5-fluorouracil and systemic pegylated interferon α-2b for advanced hepatocellular carcinoma.

400 Background: The prognosis of advanced hepatocellular carcinoma (HCC) remains poor, particularly among patients with portal vein tumor thrombosis (PVTT). This study evaluated the efficacy of combined hepatic arterial infusion chemotherapy of 5-fluorouracil (5-FU) and systemic pegylated interferon (PEG-IFN) α-2b in patients with advanced HCC. Methods: Subjects comprised 59 HCC patients with PVTT treated using subcutaneous administration of PEG-IFNα-2b (50-100 mg on day 1 of every week, for 4 weeks) and intra-arterial infusion of 5-FU (250 mg/day for 5 h on days 1-5 of every week, for 4 weeks). One treatment cycle lasted 4 weeks. Response to treatment was evaluated according to the Response Evaluation Criteria in Solid Tumors. The current therapy was discontinued in patients with progressive disease (PD). For responses other than PD, treatment was repeated for ³1 cycle. The primary efficacy endpoint was the objective early response rate. Secondary efficacy endpoints were progression-free survival (PFS) and overall survival (OS). Results: Objective early response rate was 73.0%. Cumulative PFS rates were 67.4% at 6 months, 30.2% at 12 months, 25.9% at 18 months and 20.7% at 24 months. Median PFS was 9.7 months. Cumulative survival rates were 82.4% at 6 months, 73.6% at 12 months, 52.8% at 24 months, and 44.0% at 36 months. Median survival time was 29.9 months. Prognostic factors affecting patient survival were analyzed by examining 21 potential parameters. Univariate analysis revealed two significant prognostic factors related to survival: PVTT grade (P=0.038); and therapeutic effect (P&lt;0.001). All adverse reactions were controllable by temporary suspension of treatment. Serious complications and treatment-related deaths were not observed. Conclusions: Although a prospective randomized controlled trial using a larger population of patients with advanced HCC is needed to evaluate combination therapy with 5-FU and PEG-IFNα-2b, this new combination therapy may be useful for patients with advanced HCC.

G2 checkpoint abrogator abates the antagonistic interaction between antimicrotubule drugs and radiation therapy

Background and purposeWe previously demonstrated that radiation may arrest tumor cells at G2 phase, which in turn prevents the cytotoxicity of antimicrotubule drugs and results in antagonistic interaction between these two modalities. Herein we tested whether G2 abrogators would attenuate the above antagonistic interaction and improve the therapeutic efficacy of combination therapy between radiation and antimicrotubule drugs. Materials and methodsBreast cancer BCap37 and epidermoid carcinoma KB cell lines were administered with radiation, UCN-01 (a model drug of G2 abrogator), paclitaxel or vincristine, alone or in combinations. The antitumor activities of single and combined treatments were analyzed by a series of cytotoxic, apoptotic, cell cycle, morphological and biochemical assays. ResultsUCN-01 significantly enhanced the cytotoxicity of radiation, antimitotic drugs, and their combined treatments in vitro. Further investigations demonstrated that UCN-01 attenuated radiation-induced G2 arrest, and subsequently repressed the inhibitory effect of radiation on drug-induced mitotic arrest and apoptosis. ConclusionsThis is the first report demonstrating that G2 checkpoint abrogation represses the inhibitory effect of radiation on antimicrotubule drugs, which may be implicated in cancer combination therapy. Considering that G2 abrogators are under extensive evaluation for cancer treatment, our findings provide valuable information for this class of promising compounds.

Therapeutic efficacy of combination therapy with intra‐arterial 5‐fluorouracil and systemic pegylated interferon α‐2b for advanced hepatocellular carcinoma with portal venous invasion

The prognosis of advanced hepatocellular carcinoma (HCC) remains poor, particularly among patients with portal vein tumor thrombosis (PVTT). This study evaluated the efficacy of combined 5-fluorouracil and pegylated interferon (PEG-IFN) α-2b in patients with advanced HCC. Subjects comprised 59 HCC patients with PVTT treated using subcutaneous administration of PEG-IFNα-2b (50-100 μg on day 1 of each week for 4 weeks) and intra-arterial infusion of 5-fluorouracil (250 mg/d for 5 hours on days 1-5 of each week for 4 weeks). One treatment cycle lasted 4 weeks. The current therapy was discontinued in patients with progressive disease (PD). For responses other than PD, treatment was repeated for ≥1 cycle. The primary efficacy endpoint was the objective early response rate. Secondary efficacy endpoints were progression-free survival (PFS) and overall survival. Objective early response rate was 73.0%. Cumulative PFS rates were 67.4% at 6 months, 30.2% at 12 months, 25.9% at 18 months, and 20.7% at 24 months. Median PFS was 9.7 months. Cumulative survival rates were 82.4% at 6 months, 73.6% at 12 months, 52.8% at 24 months, and 44.0% at 36 months. Median survival time was 29.9 months. All adverse reactions were controllable by temporary suspension of treatment. Serious complications and treatment-related deaths were not observed. Although a prospective randomized controlled trial using a larger population of patients with advanced HCC is needed to evaluate combination therapy with 5-fluorouracil and PEG-IFNα-2b, this new combination therapy may be useful for patients with advanced HCC.

Escherichia coli Lipopolysaccharide-Induced Immunomodulation Along With Oxytocin Administration After Mating as a Treatment Protocol for Persistent Endometritis in Mares

Endometritis is accepted as a major hindrance to achieve optimal reproductive efficiency in mares. The objective of the present study was to evaluate the therapeutic efficacy of combined therapy of immunomodulator and ecbolic as an alternative stand-alone therapy for mares with persistent endometritis. On the basis of history, culture, endometrial cytology, and per rectal and/or ultrasonographic genital examinations, 76 subfertile mares were selected and assigned to three age groups and four treatment (G-1, 2, 3) and control (G-4) groups. At estrus, all the mares were bred once naturally. Thereafter, the mares of G-1 (n = 28) were aseptically treated at 6-hours after natural service with intrauterine infusion (in 50 mL normal saline solution) of 100 μg of Escherichia coli lipopolysaccharide (LPS). Additionally, these mares received two injections of 20 IU of oxytocin (IV) at 12 and 24-hours after infusion. Mares in group G-2 (n = 11) were treated with LPS as mares of G-1, whereas mares in G-3 (n = 12) received oxytocin injections only. The mares of G-4 (n = 23) did not receive any treatment. Pregnancy rates at day 21 and foaling rates were higher ( P < .001) in group G-1 than in G-4. In G-1, higher percentage of mares at ages 6–10 years conceived and foaled than mares aged ≥16 years. On re-swabbing of mares that remained nonpregnant, the majority of G-1 and G-2 mares demonstrated sterile cultures and negative cytology, whereas uterine inflammation persisted in mares of G-3 and G-4. In conclusion, the combined therapy was effective for the elimination of persistent endometritis and improved reproductive performance of subfertile mares.

Poster 216: Combination of CTLs (CD8+/IFN) Reacted With Tumor Cells and Anti-CD25 mAb Provides Improved Immunotherapy for Tumors: An Animal Study

Immuno-therapeutic techniques for cancer were dramatic progress year by year. However, it was limited in efficacy of those. As a reason for it, regulatory T cell (Treg, CD4+/CD25+), which cells depress anti-tumor activity of T-cells in patients with cancer, was detected. On the other hand, it was observed that CTLs (CD8+) produced IFN when CTLs recognized tumor cells. In this study, we investigate therapeutic efficacy of combination therapy using CTLs (CD8+/IFN) and anti-CD25 mAb in the tumor-bearing immunocompetent animals. The RenCa (murine renal carcinoma line originated from Balb/c) xenografted mice were used as animal model. Anti-CD25 IL-2Ra mAb from hybridoma PC61 was used to deplete CD25+ cells. In vivo depletion was performed using a modification of the procedure of Onizuka et al. Mice received 250 μg of anti-CD25 mAb inoculated intraperitoneally. Tumor-bearing mice were divided into 4 administration groups: (1) whole spleen lymphocyte from tumor-bearing mice, (2) CTLs (CD8+/IFN) separated from spleen cells of tumor-bearing mice, (3) anti-CD25 mAb, (4) anti-CD25 mAb and CTLs. In group (4), CTLs were administrated into tumor-bearing mice at 1 week after treatment of anti-CD25 mAb. CTLs (CD8+/IFN) were separated from spleen cells of tumor-bearing mice using sorting system of BD FACS Vantage SE. All of mice in group (1) and (2) were death until 4 weeks after treatment in the same as untreated control mice. In group (3), tumors were disappeared in all mice at 2 days after treatment, but re-growing were observed at 5 weeks after treatment. In contrast, in group (4), tumors were disappeared in all mice at 2 days after treatment and not observed re-growing at 5 weeks after treatment. A combination of the Treg-depletion using anti-CD25 mAb and CTL administration is a feasible approach in the treatment of cancer and should be further explored in the clinic.