Efficacy Of Therapeutic Regimens Research Articles

Modeling Drug Responses and Evolutionary Dynamics using Patient-Derived Xenografts Reveals Precision Medicine Strategies for Triple Negative Breast Cancer.

The inter- and intra-tumor heterogeneity of triple negative breast cancers (TNBC), which is reflected in diverse drug responses, interplays with tumor evolution. Here, we developed a preclinical experimental and analytical framework using treatment-naive TNBC patient-derived tumor xenografts (PDTX) to test their predictive value in personalized cancer treatment approaches. Patients and their matched PDTX exhibited concordant drug responses to neoadjuvant therapy using two trial designs and dosing schedules. This platform enabled analysis of non-genetic mechanisms involved in relapse dynamics. Treatment resulted in permanent phenotypic changes with functional and therapeutic consequences. High throughput drug screening methods in ex vivo patient derived tumor xenograft cells (PDTCs) revealed patient-specific drug response changes dependent on first-line therapy. This was validated in vivo, as exemplified by a change in olaparib sensitivity in tumors previously treated with clinically relevant cycles of standard-of-care chemotherapy. In summary, PDTXs provide a robust tool to test patient drug responses and therapeutic regimens and to model evolutionary trajectories. However, high inter-model variability and permanent non-genomic transcriptional changes constrain their use for personalized cancer therapy. This work highlights important considerations associated with preclinical drug response modeling and potential uses of the platform to identify efficacious and preferential sequential therapeutic regimens.

Unraveling the Secrets Behind the Multidrug-Resistant Tuberculosis Treatment Outcome in Chronic Renal Failure Patients Requiring Hemodialysis: A Systematic Review.

Multidrug-resistant/rifampicin-resistant tuberculosis (MDR/RR TB) is a global concern, with 450,000 new cases and 191,000 deaths in 2021. TB and chronic kidney disease (CKD) have been associated since 1974, with suggested explanations such as oxidative stress, malnutrition, dysfunction in vitamin D metabolism, and a compromised cell-mediated immune response. End-stage renal failure patients are more likely to acquire drug resistance due to poor adherence, adverse drug reactions, and inappropriate dose adjustment. We then aim to evaluate the therapeutic outcome of multidrug-resistant TB of the lungs in patients who require hemodialysis in terms of successful treatment (cured and treatment completed) and the associated factors for a favorable outcome. Our secondary goal is to identify unfavorable treatment outcomes (treatment failed, patient died, or patient lost to follow-up) and the underlying associated factors. We conformed to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) 2020 Guidelines for this systematic review. We included adults (>19 years old) with chronic kidney disease who needed hemodialysis and had microbiologically confirmed multidrug-resistant pulmonary TB, excluding patients who had a renal allograft transplant, were on peritoneal dialysis, hadextrapulmonary TB, were childrenand pregnant patients. We searched PubMed, MEDLINE, PubMed Central, ScienceDirect, Public Library of Science (PLOS), and Google Scholar. Keywords were combined with the Boolean "AND" operator to gather results as well as the medical subject heading (MeSH) search strategy. After screening study articles by reading their titles and abstracts, the following tools were used to assess the risk of bias: the Newcastle-Ottawa scale for observational studies, the Assessment of Multiple Systematic Reviews (AMSTAR) checklist for systematic reviews, and the Joanna Briggs Institute (JBI) assessment tool for case reports.Primary and secondary outcomes were assessed, and a conclusion was made. We gathered 21,570 studies from the databases between 2013 and 2023, with 30,062 total participants. There were eight eligible studies for review. Patients with CKD, particularly those on dialysis, are at increased risk of TB due to a combination of factors that contribute to immunosuppression. TB reactivation is common in chronic renal failure patients. Diagnostic samples such as sputum and pleural fluid had lower sensitivity rates compared to tissue samples, which led to delays in diagnosis and treatment and, most importantly, contributed to drug resistance. All new dialysis patients should undergo interferon-gamma release assay testing. TB-infected patients with severe renal disease (eGFR 30 ml/min) had increased morbidity and mortality; however, the use of directly observed treatment, short-course (DOTS), and renal-dose adjustment of anti-TB medications significantly reduced these risks. Drug-induced hepatitis and cutaneous reactions were common adverse effects of anti-TB medications. A therapeutic drug monitoring guideline is required to reduce these adverse events and even mortality. Additional research is required to assess the safety and efficacy of therapeutic regimens, as well as their outcomes, in this population with multidrug-resistant TB.

Natural Products Lysobactin and Sorangicin A Show In Vitro Activity against Mycobacterium abscessus Complex.

The prevalence of lung disease caused by Mycobacterium abscessus is increasing among patients with cystic fibrosis. M. abscessus is a multidrug resistant opportunistic pathogen that is notoriously difficult to treat due to a lack of efficacious therapeutic regimens. Currently, there are no standard regimens, and treatment guidelines are based empirically on drug susceptibility testing. Thus, novel antibiotics are required. Natural products represent a vast pool of biologically active compounds that have a history of being a good source of antibiotics. Here, we screened a library of 517 natural products purified from fermentations of various bacteria, fungi, and plants against M. abscessus ATCC 19977. Lysobactin and sorangicin A were active against the M. abscessus complex and drug resistant clinical isolates. These natural products merit further consideration to be included in the M. abscessus drug pipeline. IMPORTANCE The many thousands of people living with cystic fibrosis are at a greater risk of developing a chronic lung infection caused by Mycobacterium abscessus. Since M. abscessus is clinically resistant to most anti-TB drugs available, treatment options are limited to macrolides. Despite macrolide-based therapies, cure rates for M. abscessus lung infections are 50%. Using an in-house library of curated natural products, we identified lysobactin and sorangicin A as novel scaffolds for the future development of antimicrobials for patients with M. abscessus infections.

Updates in the Use of BCL-2-Family Small Molecule Inhibitors for the Treatment of Relapsed/Refractory Multiple Myeloma.

Simple SummaryMonumental therapeutic advances have been made over the past two decades for the treatment of multiple myeloma. Anti-apoptotic proteins, such as Bcl-2, Bcl-xL, and Mcl-1, have been found to be upregulated in multiple myeloma cell lines, and small molecule inhibitors that target these proteins are in clinical development with the goal of enhancing apoptosis, reversing drug resistance, and improving the survival outcomes of patients with relapsed/refractory multiple myeloma. In this paper, we review the available clinical data for the Bcl-2-family protein inhibitors currently in clinical development for relapsed/refractory multiple myeloma.Despite considerable advances in the treatment of multiple myeloma over the past decade, progression of disease is inevitable, and patients ultimately succumb to relapsed and refractory disease. Efficacious therapeutic regimens that target the key biological pathways that are essential for malignant plasma cell survival are necessary in the efforts to improve patient survival outcomes. The Bcl-2 family of proteins comprise oncogenes that promote myeloma cell survival by conferring resistance to apoptosis. These proteins are frequently upregulated in myeloma cells, thus making them attractive therapeutic targets. Several small molecule inhibitors of Bcl-2-family proteins are currently in clinical development for the treatment of relapsed/refractory multiple myeloma. Venetoclax, a Bcl-2-specific inhibitor, has generated the most clinical data and has shown promising results in patients with multiple myeloma harboring the t (11;14) translocation. Venetoclax has shown efficacy when combined with anti-CD38 monoclonal antibodies, immunomodulatory drugs, and proteasome inhibitors. Several other Bcl-2 inhibitors are in clinical development, as are inhibitors of Mcl-1, a Bcl-2-family oncoprotein that is perhaps more critical for myeloma cell survival than Bcl-2. This review will summarize the latest clinical data regarding the clinical development of Bcl-2-family protein inhibitors in the treatment of relapsed/refractory multiple myeloma.

Per a 5-derived T-cell peptides modulate NF-kB signalling to ameliorate allergic inflammation systemically in murine model of cockroach allergic hyper-reactivity.

Peptide immunotherapy (PIT) represents a safe and efficacious therapeutic regimen with in-consequential side-effects. The present study aims to identify T-cell epitopes of Per a 5 allergen, a delta class GST from Periplaneta americana and investigate effect of peptide treatment in murine model of cockroach allergen-mediated hyper-reactivity. The epitopes (TC-P1, TC-P2, and TC-P3) were identified as promiscuous MHC-II binders by MHC-Pred, ProPred, and IEDB analysis tool. Murine model of cockroach allergic hyper-reactivity was generated in Balb/c mice. A marked reduction in cellular infiltration in lungs (3-fold compared with Non-IT) was observed in T3-IT group as evidenced by total leucocyte count in BALF and histology. Specific IgE levels were reduced 3-fold in T2-IT and T3-IT compared with Non-IT with increase in IgG2a levels. IL-4 and IL-13 were reduced upto 2.5-fold in treatment groups compared with Non-IT group. Splenocytes revealed significant increase in levels of CD4+FoxP3+ T cells in TC-P1 and TC-P2 mice demonstrating a systemic shift towards Tregs. Peptide treatment downregulated NF-kB signalling in lung and enhanced the levels of immune-regulatory molecules α1-antitrypsin and elafin. Our results indicate that TC-P1 and TC-P3 alter Th2 cytokine milieu and antibody isotype ratio to suppress allergic inflammation. PIT modulates local and systemic mechanisms to resolve inflammation and possess potential for treatment in cockroach allergy.

Epigenetic enzyme mutations as mediators of anti-cancer drug resistance.

Despite the rapid advancement in the introduction of new drugs for cancer therapy, the frequent emergence of drug resistance leads to disease progression or tumor recurrence resulting in dismal prognosis. Given that genetic mutations are thought to be important drivers of anti-cancer drug resistance, it is of paramount importance to pin-point mutant genes that mediate drug resistance and elucidate the underlying molecular mechanisms in order to develop novel modalities to surmount chemoresistance and achieve more efficacious and durable cancer therapies. Cumulative evidence suggests that epigenetic alterations, especially those mediated by epigenetic enzymes with high mutation rates in cancer patients, can be a crucial factor in the development of chemoresistance. Mutant epigenetic enzymes have altered enzymatic activity which may directly or indirectly affect the level of histone modifications. This can change chromatin structure and function hence altering the expression of target genes and eventually lead to chemoresistance. In the current review, we summarize epigenetic enzyme mutations and the consequent mechanisms of drug resistance in pre-clinical drug-resistance models and relapsed cancer patient specimens. We also introduce previously unreported mutation sites in the DOT1 domain of DOT1L, which are related to lung cancer drug resistance. It is worth noting that mutations occur not only in domains with enzymatic activity but also in non-catalytic regions. Each protein domain is an evolutionarily conserved region with independent functional properties. This may provide a rationale for the potential development of small molecule inhibitors which target various functional domains of epigenetic enzymes. Finally, based on the multitude of mechanisms of drug resistance, we propose several therapeutic strategies to reverse or overcome drug-resistance phenotypes, with the aim to provide cancer patients with novel efficacious combination therapeutic regimens and strategies to improve patient prognosis.

P87.03 Characteristics and Outcomes of RET-Rearranged Non-Small Cell Lung Cancer from China: A Retrospective Study of Real-World

Rearranged during transfection (RET) proto-oncogene gene fusions are rare in non–small-cell lung cancer (NSCLC), about 1-2%. RET-rearranged tumours have been shown to respond to tyrosine kinase inhibitors, such as cabozantinib, vandetanib. Several clinical trials also has been undergoing. In the real word, RET fusion patients characteristics, treatment options and outcomes are still not clear. A retrospective, single center study of patients with pathologically confirmed NSCLC was conducted. RET rearrangements were detected using next generation sequencing (NGS). We analyzed the clinical characteristics of patients with RET-rearranged NSCLC and the efficacy of therapeutic regimens. We also evaluated the RET-rearranged patients survival outcomes, PFS of first-line therapy (PFS1), and second line therapy (PFS2). From 9/July/ 2017 to 25/Nov/2019, Total 3217 NSCLC patients were screened for RET-rearranged testing, only 41 (1.3%) patients had RET fusion gene, including 23 KIF5B-RET,10 CCDC6-RET and 8 other RET fusion gene. The average age was 60 years old (30-85). Of the 41 patients, 23(56%) females and 18 males, 36(87.8%) never smoker, 5 heavy smokers and no light smokers, 39(95%) adenocarcinoma and 2 squamous carcinoma. Most (33/41) of them had advanced stage (IIIB/IIIC/IV) . the majority (35/41) of patients had lymph node metastasis, especially in mediastinum, neck, supraclavicular, and axillary regions. Significantly, the axillary lymph node metastasis reached 12% (4/33). The common distant metastasis was still brain, bone, liver and adrenal gland. Unfortunately, 9 patients had no treatment records and failed to follow up. 20 patients with stage IIIB to stage IV had accepted first-line treatment, including 17chemotherapy, 2 target therapy, and 1 immunotherapy. The median progression-free survival (PFS1) of first-line chemotherapy was 9.3m (7.0m-11.6m), 2 patients with cabozantinib PFS1 were 3.9m and 6.0m respectively. the only one with duravalumab was 6.0m. There were few patients accepted second line therapy, The PFS2 was 3.0m(1.6m-6.5m). The median OS of all advanced (stage IIIB/IV) patients was 12.0 (6.5m-17.5m). Further analysis found that there were 20 patients accepted one or more lines treatment, while 4 patients had died through non-treatment. The OS of treated and non-treated advanced NSCLC patients were 14.0m and 2.0m respectively.Table 1RET-rearranged NSCLC characteristic and therapeutic outcometotal (n=41)N(%)Treatment outcomeAge,years60(30-85)First-linePFS1Sexchemotherapy9.3 (7.0m-11.6m)female23 (56.0%)target therapy3.9m/6.0mmale18 (44.0%)immunotherapy6.0mSmokeSecond-linePFS2never36 (87.8%)chemotherapy3.0m (1.4m-8.5m)light0target therapy3.0m (1.0m-3.6m)heavy5 (12.2%)OSRET-rearranged siteAll patients12.0m (6.5m-17.5m)KIF5B-RET23 (56.1%)treated14.0m (1.7m-26.2m)CCDC6-RET10 (24.4%)non-treated2.0m (0.4m-3.6m)others8 (19.5%)Pathology typeadenocarcinoma38 (92.7%)sqmous carcinoma2 (4.9%)neuroendocrine tumor1 (2.4%)Lymph node metastasismediastinum20 (47.8%)supraclavicular15 (36.6%)axillary4 (9.8%) Open table in a new tab RET-rearranged NSCLC was more likely to have lymph node metastasis. Because of poor access to target drug of cabozantinib and vandetanib in China, most of RET rearranged non-small cell lung cancer had chosen chemotherapy as first-line therapy, even so, they had a quite acceptable PFS, and overall survival was prolonged.

Mediator subunit MED1 is required for E2A-PBX1–mediated oncogenic transcription and leukemic cell growth

The chimeric transcription factor E2A-PBX1, containing the N-terminal activation domains of E2A fused to the C-terminal DNA-binding domain of PBX1, results in 5% of pediatric acute lymphoblastic leukemias (ALL). We recently have reported a mechanism for RUNX1-dependent recruitment of E2A-PBX1 to chromatin in pre-B leukemic cells; but the subsequent E2A-PBX1 functions through various coactivators and the general transcriptional machinery remain unclear. The Mediator complex plays a critical role in cell-specific gene activation by serving as a key coactivator for gene-specific transcription factors that facilitates their function through the RNA polymerase II transcriptional machinery, but whether Mediator contributes to aberrant expression of E2A-PBX1 target genes remains largely unexplored. Here we show that Mediator interacts directly with E2A-PBX1 through an interaction of the MED1 subunit with an E2A activation domain. Results of MED1 depletion by CRISPR/Cas9 further indicate that MED1 is specifically required for E2A-PBX1-dependent gene activation and leukemic cell growth. Integrated transcriptome and cistrome analyses identify pre-B cell receptor and cell cycle regulatory genes as direct cotargets of MED1 and E2A-PBX1. Notably, complementary biochemical analyses also demonstrate that recruitment of E2A-PBX1 to a target DNA template involves a direct interaction with DNA-bound RUNX1 that can be further stabilized by EBF1. These findings suggest that E2A-PBX1 interactions with RUNX1 and MED1/Mediator are of functional importance for both gene-specific transcriptional activation and maintenance of E2A-PBX1-driven leukemia. The MED1 dependency for E2A-PBX1-mediated gene activation and leukemogenesis may provide a potential therapeutic opportunity by targeting MED1 in E2A-PBX1+ pre-B leukemia.

Leveraging Patient-Derived Models for Immunotherapy Research.

As cancer immunotherapies become mainstream for the treatment of many different cancer types and the numbers of new agents continue to increase, the need for experimental models is also rising. An approach to develop and study models for immune-oncology that has garnered intense interest in recent years is that of using patient-derived models. Patient-derived models can recapitulate many of the features and heterogeneity of the corresponding human tumors. Historically these models have been used to study cancer cell-intrinsic properties of tumors and drugs that target tumor cells directly. In recent years, increasing recognition of the role immune cells play in cancer and how these represent good therapeutic targets has led to efforts to optimize and use patient-derived models for cancer immunotherapy studies. Patient-derived models are now being used to study the properties of cancer cells that modulate their ability to respond to immune stimulation. Further efforts are underway to use and develop patient-derived models that incorporate human immune cells in vitro and in vivo (humanized mice) so that cancer cell-immune cell interactions can be studied in the context of cancer immunotherapies. As these models are further refined, leveraging patient-derived models for cancer immunotherapy research can provide insight into mechanisms of sensitivity and resistance to cancer immunotherapies, uncover new targets, reveal how specific agents work, and be used to evaluate the antitumor efficacy of therapeutic regimens.

Advanced technological tools to study multidrug resistance in cancer.

The complexity of cancer biology and its clinical manifestation are driven by genetic, epigenetic, transcriptomic, proteomic and metabolomic alterations, supported by genomic instability as well as by environmental conditions and lifestyle factors. Although novel therapeutic modalities are being introduced, efficacious cancer therapy is not achieved due to the frequent emergence of distinct mechanisms of multidrug resistance (MDR). Advanced technologies with the potential to identify and characterize cancer MDR could aid in selecting the most efficacious therapeutic regimens and prevent inappropriate treatments of cancer patients. Herein, we aim to present technological tools that will enhance our ability to surmount drug resistance in cancer in the upcoming decade. Some of these tools are already in practice such as next-generation sequencing. Identification of genes and different types of RNAs contributing to the MDR phenotype, as well as their molecular targets, are of paramount importance for the development of new therapeutic strategies aimed to enhance drug response in resistant tumors. Other techniques known for many decades are in the process of adaptation and improvement to study cancer cells' characteristics and biological behavior including atomic force microscopy (AFM) and live-cell imaging. AFM can monitor in real-time single molecules or molecular complexes as well as structural alterations occurring in cancer cells induced upon treatment with various antitumor agents. Cell tracking methodologies and software tools recently progressed towards quantitative analysis of the spatio-temporal dynamics of heterogeneous cancer cell populations and enabled direct monitoring of cells and their descendants in 3D cultures. Besides, novel 3D systems with the advanced mimicking of the in vivo tumor microenvironment are applicable to study different cancer biology phenotypes, particularly drug-resistant and aggressive ones. They are also suitable for investigating new anticancer treatment modalities. The ultimate goal of using phenotype-driven 3D cultures for the investigation of patient biopsies as the most appropriate in vivo mimicking model, can be achieved in the near future.

Pharmacological and non-pharmacological treatments for opioid-induced constipation: A protocol for systematic review and network meta-analysis.

Background:Pain is very common and its management with a huge burden for patients and the healthcare system. And the network meta-analysis was designed to provide reference for the clinical practice.Methods:PubMed, EMBASE, Cochrane library, CNKI, VIP, Wan Fang, and CBM will be systematically searched their inception to November 2018. Randomized controlled trials that compared the effect of differently pharmacological or non-pharmacological treatments for opioid-induced constipation will be included. The primary outcome is the efficacy of therapeutic regimens. Risk of bias assessment of the included studies will be performed using the Cochrane risk of bias tool. A network meta-analysis will be performed using STATA 13.0 software with WinBUGS 1.4.3 software. Grading of Recommendations Assessment, Development, and Evaluation will be used to assess the overall quality of evidence.Results:This study is ongoing and will be submitted to a peer-reviewed journal for publication.Conclusion:This study will provide a comprehensive evidence on the effectiveness and safety of pharmacological and non-pharmacological treatments for opioid-induced constipation.PROSPERO registration number:CRD42018116533.

Toward fully exploiting the therapeutic potential of marketed pharmaceuticals: the use of octreotide and chloroquine in oncology.

Pleiotropy in biological systems and their targeting allows many pharmaceuticals to be used for multiple therapeutic purposes. Fully exploiting the therapeutic properties of drugs that are already marketed would be highly advantageous. This is especially the case in the field of oncology, where the ineffectiveness of typical anticancer agents is a common issue, while the development of novel anticancer agents is a costly and particularly time-consuming process. Octreotide and chloroquine are two pharmaceuticals that exhibit profound antitumorigenic activities. However, the current therapeutic use of octreotide is restricted primarily to the management of acromegaly and neuroendocrine tumors, both of which are rare medical conditions. Similarly, chloroquine is used mainly for the treatment of malaria, which is designated as a rare disease in Western countries. This limited exploitation contradicts the experimental findings of numerous studies outlining the possible expansion of the use of octreotide to include the treatment of common human malignancies and the repositioning of chloroquine in oncology. Herein, we review the current knowledge on the antitumor function of these two agents stemming from preclinical or clinical experimentation. In addition, we present in silico evidence on octreotide potentially binding to multiple Wnt-pathway components. This will hopefully aid in the design of new efficacious anticancer therapeutic regimens with minimal toxicity, which represents an enormous unmet demand in oncology.

The impact of location on the prognosis of squamous cell carcinomas of the anorectal region

BackgroundBecause anal and rectal squamous cell carcinomas (R-SCCs and A-SCCs) share a common histology and an excellent response to chemoradiation, we hypothesized that R-SCC and A-SCC may represent a similar biological entity, and location would not affect clinical presentation and prognosis. MethodsPatients diagnosed with R-SCC (n = 2881) and A-SCC (n = 21,854) were identified in the Surveillance, Epidemiology, and End Results database (1998-2013). R-SCCs were staged based on American Joint Committee on Cancer classification for A-SCC, and impact of location was analyzed accordingly. ResultsCompared to A-SCC, R-SCCs were more common in females (65% versus 48%, P < 0.001) and older patients (62 versus 56 yrs, P < 0.001). R-SCC presented with more advanced disease than A-SCC: mean size 4.2 versus 3.6 cm; T4 14% versus 5%; nodal involvement 20% versus 15%; and metastases 13% versus 6% (all P < 0.001). In multivariable analysis, R-SCCs and A-SCCs had similar disease-specific survival (DSS) for stages 0, I, and III; however, stage II R-SCC had significantly worse DSS than A-SCCs (P = 0.002). This was due to a greater proportion of T3 (>5 cm) R-SCC tumors (36% versus 27%, P < 0.001), which had a lower DSS than T2 (2-5 cm) tumors. Within T3 and T4 tumors, R-SCCs had lower DSS than A-SCCs. ConclusionsR-SCC presented with higher stages than A-SCC, suggesting a delayed diagnosis. Larger R-SCC (T3-T4) had worse survival compared to T3-4 A-SCC, which may be due to a combination of more advanced disease within-stage as well as the use of less efficacious therapeutic regimens. Therefore, location may represent a significant prognostic factor for SCC of the anorectal region.

Progesterone action in breast, uterine, and ovarian cancers.

Progesterone and progesterone receptors (PRs) are essential for the development and cyclical regulation of hormone-responsive tissues including the breast and reproductive tract. Altered functions of PR isoforms contribute to the pathogenesis of tumors that arise in these tissues. In the breast, progesterone acts in concert with estrogen to promote proliferative and pro-survival gene programs. In sharp contrast, progesterone inhibits estrogen-driven growth in the uterus and protects the ovary from neoplastic transformation. Progesterone-dependent actions and associated biology in diverse tissues and tumors are mediated by two PR isoforms, PR-A and PR-B. These isoforms are subject to altered transcriptional activity or expression levels, differential crosstalk with growth factor signaling pathways, and distinct post-translational modifications and cofactor-binding partners. Herein, we summarize and discuss the recent literature focused on progesterone and PR isoform-specific actions in breast, uterine, and ovarian cancers. Understanding the complexity of context-dependent PR actions in these tissues is critical to developing new models that will allow us to advance our knowledge base with the goal of revealing novel and efficacious therapeutic regimens for these hormone-responsive diseases.

Assessing the therapeutic efficacy of oxime therapies against percutaneous organophosphorus pesticide and nerve agent challenges in the Hartley guinea pig.

Given the rapid onset of symptoms from intoxication by organophosphate (OP) compounds, a quick-acting, efficacious therapeutic regimen is needed. A primary component of anti-OP therapy is an oxime reactivator to rescue OP-inhibited acetylcholinesterases. Male guinea pigs, clipped of hair, received neat applications of either VR, VX, parathion, or phorate oxon (PHO) at the 85(th) percentile lethal dose, and, beginning with presentation of toxicosis, received the human equivalent dose therapy by intramuscular injection with two additional follow-on treatments at 3-hr intervals. Each therapy consisted of atropine free base at 0.4 mg/kg followed by one of eight candidate oximes. Lethality rates were obtained at 24 hr after VR, VX and PHO challenges, and at 48 hr after challenge with parathion. Lethality rates among symptomatic, oxime-treated groups were compared with that of positive control (OP-challenged and atropine-only treated) guinea pigs composited across the test days. Significant (p ≤ 0.05) protective therapy was afforded by 1,1-methylene bis(4(hydroxyimino- methyl)pyridinium) dimethanesulfonate (MMB4 DMS) against challenges of VR (p ≤ 0.001) and VX (p ≤ 0.05). Lethal effects of VX were also significantly (p ≤ 0.05) mitigated by treatments with oxo-[[1-[[4-(oxoazaniumylmethylidene)pyridin-1-yl]methoxymethyl]pyridin-4-ylidene]methyl]azanium dichloride (obidoxime Cl2) and 1-(((4-(aminocarbonyl) pyridinio)methoxy)methyl)-2,4-bis((hydroxyimino)methyl)pyridinium dimethanesulfonate (HLö-7 DMS). Against parathion, significant protective therapy was afforded by obidoxime dichloride (p ≤ 0.001) and 1,1'-propane-1,3-diylbis{4-[(E)-(hydroxyimino)methyl]pyridinium} dibromide (TMB-4, p ≤ 0.01). None of the oximes evaluated was therapeutically effective against PHO. Across the spectrum of OP chemicals tested, the oximes that offered the highest level of therapy were MMB4 DMS and obidoxime dichloride.

Progranulin promotes Temozolomide resistance of glioblastoma by orchestrating DNA repair and tumor stemness.

Glioblastoma multiforme (GBM) is the most common malignant brain tumor in adults with a dismal prognosis. Current therapy of surgical removal combined with Temozolomide (TMZ) and radiation therapy only slightly prolongs the survival of GBM patients. Thus, it is essential to elucidate mechanism underlying its highly malignant properties in order to develop efficacious therapeutic regimens. In this study, we showed that progranulin (PGRN) was overexpressed in most GBM cell lines and the majority of human tumor samples. PGRN overexpression conferred GBM cells with tumorigenic properties and TMZ resistance by upregulating DNA repair (PARP, ATM, BRCA1, Rad51, XRCC1 and so on) and cancer stemness (CD133, CD44, ABCG2) genes, in part via an AP-1 transcription factor, specifically cFos/JunB. Curcumin, an AP-1 inhibitor, was also found to regulate PGRN promoter activity and expression including its downstream effectors aforementioned. These data suggested a feedforward loop between PGRN signaling and AP-1. PGRN depletion significantly decreased unlimited self-renewal and multilineage differentiation and the malignant properties of GBMs cells S1R1, and enhanced their vulnerability to TMZ. In addition, S1R1 depleted of PGRN also lost the ability to form tumor in an orthotopic xenograft mouse model. In conclusion, PGRN had a critical role in the pathogenesis and chemoresistance of GBM and functioned at the top of the hierarchy of cellular machinery that modulates both DNA repair pathways and cancer stemness. Our data suggest that a new strategy combining current regimens with compounds targeting PGRN/AP-1 loop like curcumin may significantly improve the therapeutic outcome of GBM.

Diffuse Alveolar Haemorrhage in ANCA-associated Vasculitis

Diffuse alveolar haemorrhage (DAH) is a serious complication of anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV). A literature review was performed to ascertain the diagnostic features, treatment, and outcome of this rare but serious condition. Haemoptysis and dyspnoea are common but non-specific features. Chest radiography is usually abnormal, and high-resolution computerised tomographic scanning is more sensitive. Increased uptake of inhaled carbon monoxide and reduced clearance of C(15)O on lung function testing is suggestive of intra-alveolar blood. Fiberoptic bronchoscopy and bronchoalveolar lavage are useful when a super-added infection is suspected. Concurrent renal disease is common and contributes to the morbidity and mortality. Treatment should be individualised, and it is based on glucocorticoid and cyclophosphamide induction with azathioprine maintenance. The role of plasmapheresis is unclear, and is currently being evaluated. Patients are at risk of disease and treatment-related long-term complications. Ongoing research into the most efficacious therapeutic regimens associated with the least side effects is especially important.

Linagliptin/Metformin Fixed-Dose Combination Treatment: A Dual Attack to Type 2 Diabetes Pathophysiology

Combination therapies are a widely accepted approach to type 2 diabetes treatment, considering that monotherapies fail to provide adequate glycemic control in the majority of cases. The combination of oral antidiabetic agents into a single tablet would significantly simplify the therapeutic regimen and maximize patients' adherence to treatment. Recently, a fixed-dose, single-tablet, combined formulation of linagliptin (a dipeptidyl peptidase-4 inhibitor) and metformin has been approved for use in type 2 diabetic patients, and is indicated as an adjunct to diet and exercise for those patients who remain inadequately controlled despite maximal tolerated doses of metformin, metformin and sulfonylurea, or linagliptin and metformin monotherapies. The combination tablet is administered twice daily and can be used either alone or combined with sulfonylureas. Clinical trials suggest that this fixed-dose combination provides significantly superior glycemic control compared to linagliptin and metformin monotherapy, in terms of improving key parameters of glucose homeostasis such as glycosylated hemoglobin, fasting and postprandial glucose levels. It also exhibits an excellent tolerability profile, without promoting weight gain and hypoglycemic episodes. The compounds of this formulation do not display clinically relevant pharmacokinetic interactions with each other, and exert synergistic (complementary) pharmacodynamic effects, including an enhanced incretin effect, suppressed hepatic glucose production, and improved peripheral insulin sensitivity. As a result, a linagliptin/metformin fixed-dose combination offers the potential to address multiple defects of type 2 diabetes pathophysiology (pancreatic islet dysfunction, insulin resistance, increased hepatic glucose output), and most importantly, in the context of a safe, efficacious, flexible, and convenient therapeutic regimen.

Abstract 1140: Intratumoral mutual exclusivity of dual amplified receptor tyrosine kinase genes in glioblastoma

Abstract Cancer heterogeneity presents a major barrier for the development of effective treatment strategies aimed at combating the disease. Within glioblastoma, this intratumoral heterogeneity may refer to the concurrent observation of multiple cellular morphologies, differential patterns of vascular proliferation, discrete transcript and/or protein expression patterns, and genetic variegation. DNA copy number profiling has demonstrated that multiple amplifications may frequently be found in the same glioblastoma specimens. Although tumour clonality would imply that these events would be present in all neoplastic cells, we and others have previously noted through detailed FISH/CISH experiments on pathological specimens that not all cells harbour individual amplification events. In order to explore these concepts more thoroughly, tissue microarrays comprising 342 high grade glioma samples were screened by FISH for cells amplified for genes encoding either EGFR or PDGFRA. A total of 18 cases (8%) contained both gene amplifications in at least one cell of the TMA core. We utilised two-colour FISH with differential labelling of probes specific to EGFR and PDGFRA on whole sections from these samples, and assessed the relative copy numbers per cell of each gene. The most common pattern comprised mutual exclusivity of gene amplification within adjacent glioblastoma cells. To quantify the extent to which these patterns were reflected across the tumour, we carefully mapped the relative copy numbers of EGFR and PDGFRA in cells from throughout the pathology specimen of all dual-amplified glioblastoma cases. By counting signals in 41,997 cells from 190 distinct loci across 17 samples, we noted a startling heterogeneity of amplification patterns both across and within tumour specimens, with approximately three-quarters of cells harbouring amplification of only one or other of the genes. In only one case did the single-cell ‘co-amplification’ model represent more than 15% of the sample as a whole. Within an individual sample, cells harbouring one, both, or neither amplification could be found in highly variable frequencies, either presenting as a mosaic of genetically distinct cells, or forming foci where one event would strongly predominate. We observed that these PDGFRA-restricted cells tended to be present close to endothelial cells, whilst poorly vascularised areas were largely the domain of EGFR-amplified tumour cells. EGFR amplification was significantly linked with areas of glioblastoma containing large numbers of giant cells and/or small cells, whereas all high frequency PDGFRA-amplified cells were found in areas of fibrillary histology. These data have profound implications for designing efficacious therapeutic regimens, as the relative contributions of cell populations harbouring one or other gene amplification to disease propagation, and the implications for targeted therapies, are not known. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1140. doi:1538-7445.AM2012-1140

Abstract 2562: Ultra sensitive immuno-array based multiplexed assay capable of profiling RTKs and signaling proteins, heterodimers, and growth factors for routine analysis of clinical samples with limited availability

Abstract Patient stratification based on target protein expressions or mutations alone has limitations in selecting efficacious therapeutic regimens for solid tumors. Information on the activation status of signaling pathway proteins as determined by their phosphorylation states and the level of expression, profiling of growth factors responsible for its activation, and their dimerization partners, can significantly influence clinical decisions. Herein we report a comprehensive analysis of key receptor tyrosine kinases (HER1, HER2, p95HER2, HER3, cMET, IGF1R, FGFRs, and others) and their downstream signal proteins (PI3K, Shc, AKT, MEK, ERK, PRAS40, RPS6, P70S6K, RSK), and dimerization partners (HER1:HER2, HER2:HER3, HER1:HER3, and others), and secreted growth factors (AREG, EREG, NRG1, HB-EGF, HGF, BTC, EGF, TGF, VEGF, and IGF-1) based on a multiplexed immunoarray CEER (Collaborative Enzyme Enhanced Reactive-immunoassay) platform. CEER platform utilizes the formation of an unique “triple-antibody-enzyme-channeling” immuno-complex. This complex requires co-localization of two detector-antibodies conjugated with corresponding channeling-enzymes once target proteins are bound by the capture antibodies. The immuno-collaboration between two detector enzymes, glucose oxidase (GO, conjugated to anti-target protein antibodies) and horseradish peroxidase (HRP, conjugated to anti-phosphorylated sites within the target) via target-specific, tyramide-mediated signal amplification enables target protein expression /phosphorylation profiling with validated analytical sensitivity between 1 to 10 cells. Using the CEER platform, quantitative modulations of target protein(s) in response to targeted agent(s) and treatment effect on associated downstream and upstream pathway proteins were measured. Due to the extreme sensitivity and specificity, CEER can be utilized in clinical samples with limited availability (e.g., FNA or CTCs). A comprehensive signaling pathway analysis can be a critical guide for selecting effective clinical strategies for targeted drug combinations or sequential therapeutic regimens. In addition, continued signaling pathway profiling after drug administration can monitor drug response and provide valuable information on potential drug resistance mechanisms. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2562. doi:1538-7445.AM2012-2562