Current Practices in Preclinical Drug Development

Schultze, A. Eric; Walker, Dana B.; Turk, James R.; Tarrant, Jacqueline M.; Brooks, Marjory B.; Pettit, Syril

doi:10.1177/0192623312460924

Cited by 28 publications

(6 citation statements)

References 21 publications

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“…miRNA expression atlases and candidate biomarkers of organ specific injury have been investigated to varying degrees in rodents and humans [ 5 , 12 , 21 ]. Dogs are a relevant preclinical species that are used in nonclinical safety assessments [ 22 , 23 ]. However, little is known about dog miRNA expression or annotation of sequences.…”

Section: Introductionmentioning

confidence: 99%

The beagle dog MicroRNA tissue atlas: identifying translatable biomarkers of organ toxicity

et al. 2016

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BackgroundMicroRNAs (miRNA) are varied in length, under 25 nucleotides, single-stranded noncoding RNA that regulate post-transcriptional gene expression via translational repression or mRNA degradation. Elevated levels of miRNAs can be detected in systemic circulation after tissue injury, suggesting that miRNAs are released following cellular damage. Because of their remarkable stability, ease of detection in biofluids, and tissue specific expression patterns, miRNAs have the potential to be specific biomarkers of organ injury. The identification of miRNA biomarkers requires a systematic approach: 1) determine the miRNA tissue expression profiles within a mammalian species via next generation sequencing; 2) identify enriched and/or specific miRNA expression within organs of toxicologic interest, and 3) in vivo validation with tissue-specific toxicants. While miRNA tissue expression has been reported in rodents and humans, little data exists on miRNA tissue expression in the dog, a relevant toxicology species. The generation and evaluation of the first dog miRNA tissue atlas is described here.ResultsAnalysis of 16 tissues from five male beagle dogs identified 106 tissue enriched miRNAs, 60 of which were highly enriched in a single organ, and thus may serve as biomarkers of organ injury. A proof of concept study in dogs dosed with hepatotoxicants evaluated a qPCR panel of 15 tissue enriched miRNAs specific to liver, heart, skeletal muscle, pancreas, testes, and brain. Dogs with elevated serum levels of miR-122 and miR-885 had a correlative increase of alanine aminotransferase, and microscopic analysis confirmed liver damage. Other non-liver enriched miRNAs included in the screening panel were unaffected. Eli Lilly authors created a complimentary Sprague Dawely rat miRNA tissue atlas and demonstrated increased pancreas enriched miRNA levels in circulation, following caerulein administration in rat and dog.ConclusionThe dog miRNA tissue atlas provides a resource for biomarker discovery and can be further mined with refinement of dog genome annotation. The 60 highly enriched tissue miRNAs identified within the dog miRNA tissue atlas could serve as diagnostic biomarkers and will require further validation by in vivo correlation to histopathology. Once validated, these tissue enriched miRNAs could be combined into a powerful qPCR screening panel to identify organ toxicity during early drug development.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-2958-x) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

The beagle dog MicroRNA tissue atlas: identifying translatable biomarkers of organ toxicity

et al. 2016

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“…Coincident with global increases in prevalence of these disorders are more frequent reports of drug-induced thrombotic events [13, 14]. Our recent survey of preclinical drug development revealed a critical gap in early detection of incipient thrombosis in the absence of lethal and grossly visible thrombi on post mortem examination in animal models [15]. In this time course study of sub-lethal LPS administration in rats, we combined comprehensive clinicopathologic tests and detailed histologic review with newer techniques analyzing extracellular vesicles and miRNA, to identify circulating biomarkers with potential applications as screening tests of procoagulant imbalance.…”

Section: Introductionmentioning

confidence: 99%

Non-Lethal Endotoxin Injection: A Rat Model of Hypercoagulability

et al. 2017

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Systemic inflammation co-activates coagulation, which unchecked culminates in a lethal syndrome of multi-organ microvascular thrombosis known as disseminated intravascular coagulation (DIC). We studied an endotoxin-induced inflammatory state in rats to identify biomarkers of hemostatic imbalance favoring hypercoagulability. Intraperitoneal injection of LPS at 15 mg/kg body weight resulted in peripheral leukopenia and widespread neutrophilic sequestration characteristic of an acute systemic inflammatory response. Early indicators of hemostatic pathway activation developed within 4 hours, including increased circulating concentrations of procoagulant extracellular vesicles (EVs), EVs expressing endothelial cell and platelet membrane markers, and high concentration of soluble intercellular adhesion molecule-1 (sICAM-1), plasminogen activator inhibitor-1 (PAI-1), and D-dimers. Inflammation persisted throughout the 48-hour observation period; however, increases were found in a subset of serum microRNA (miRNA) that coincided with gradual resolution of hemostatic protein abnormalities and reduction in EV counts. Dose-adjusted LPS treatment in rats provides a time-course model to develop biomarker profiles reflecting procoagulant imbalance and rebalance under inflammatory conditions.

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“… 18 There are risks associated with oversimplifying preclinical platelet studies and extrapolating findings using healthy donor platelets to studies with platelets from a diseased population. 19 A prudent research approach may include determining whether the signaling processes in platelets from diseased patients are similar to healthy persons. If platelets from a diseased population have different agonist signaling properties, the design and implementation of antiplatelet agents should be tailored to adjust for these changes.…”

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confidence: 99%

Hypoxia and Ischemia Promote a Maladaptive Platelet Phenotype

Cameron

Mix

Ture

et al. 2018

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Objective Reduced blood flow and/or tissue oxygen tension conditions result from thrombotic and vascular diseases such as myocardial infarction, stroke, and peripheral vascular disease. It is largely assumed that while platelet activation is increased by an acute vascular event, chronic vascular inflammation, and/or ischemia, the platelet activation pathways and responses are not themselves changed by the disease process. We therefore sought to determine whether the platelet phenotype is altered by hypoxic and ischemic conditions. Approach and Results In a cohort of patients with metabolic and peripheral artery disease (PAD), platelet activity was enhanced and/or inhibition with oral anti-platelet agents was impaired compared to platelets from control subjects, suggesting a difference in platelet phenotype caused by disease. Isolated murine and human platelets exposed to reduced oxygen (hypoxia chamber, 5% O2) had increased expression of some proteins that augment platelet activation compared to platelets in normoxic conditions (21% O2). Using a murine model of critical limb ischemia (CLI), platelet activity was increased even two weeks post-surgery compared to sham surgery mice. This effect was partly inhibited in platelet specific Extracellular Regulated Protein Kinase 5 (ERK5) knockout mice. Conclusions These findings suggest that ischemic disease changes the platelet phenotype and alters platelet agonist responses due to changes in the expression of signal transduction pathway proteins. Platelet phenotype and function should therefore be better characterized in ischemic and hypoxic diseases to understand the benefits and limitations of anti-platelet therapy.

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Current Practices in Preclinical Drug Development

Cited by 28 publications

References 21 publications

The beagle dog MicroRNA tissue atlas: identifying translatable biomarkers of organ toxicity

The beagle dog MicroRNA tissue atlas: identifying translatable biomarkers of organ toxicity

Non-Lethal Endotoxin Injection: A Rat Model of Hypercoagulability

Hypoxia and Ischemia Promote a Maladaptive Platelet Phenotype

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