RESUMO
The identification, application, and qualification of safety biomarkers are becoming increasingly critical to successful drug discovery and development as companies are striving to develop drugs for difficult targets and for novel disease indications in a risk-adverse environment. Translational safety biomarkers that are minimally invasive and monitor drug-induced toxicity during human clinical trials are urgently needed to assess whether toxicities observed in preclinical toxicology studies are relevant to humans at therapeutic doses. The interpretation of data during the biomarker qualification phase should include careful consideration of the analytic method used, the biology, pharmacokinetic and pharmacodynamic properties of the biomarker, and the pathophysiology of the process studied. The purpose of this review is to summarize commonly employed technologies in the development of fluid- and tissue-based safety biomarkers in drug discovery and development and to highlight areas of ongoing novel assay development.
Assuntos
Biomarcadores Farmacológicos/análise , Descoberta de Drogas/métodos , Avaliação Pré-Clínica de Medicamentos , Animais , Ensaios Clínicos como Assunto , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Humanos , Patologia Clínica , Pesquisa Translacional BiomédicaRESUMO
This study directly demonstrates that cardiac troponin I (cTnI) is a sensitive, specific, and persistent biomarker in laboratory animals. Histopathological and pathophysiological cardiac changes in dogs, rats and mice correlated with increased serum cTnI with various cardiac inotropic agents, and cardiotoxic drugs and with cardiac arrhythmias, tachycardia, cardiac effusion with dyspnoea, and ageing. A comparison of six immunoassays for cTnI and cardiac troponin T (cTnT) to detect and monitor cardiac injury in a rodent model indicated that enzyme-linked immunosorbent (Life Diagnostics Inc and TriChem Resources Inc, West Chester, Philadelphia, USA) and Immulite (Diagnostic Products Corporation, Llanberis, UK) assays had low sensitivity and less than 1% of the dynamic range of Centaur (Bayer Healthcare Diagnostics, Newbury, UK) cTnI and Elecsys (Roche Diagnostics, Basel, Switzerland) and M8 (Bioveris Europe, Whitney, UK) cTnT assays. In dogs, however, the Immulite assay was effective and correlated with the Centaur. Serum concentrations were highly correlated but 10-fold lower for cTnT compared with cTnI with cardiac injury. Centaur assay also detected cTnI in myocardium from marmosets, swine, cattle, and guinea pigs, indicating it to be candidate cardiac biomarker for these species as well. Purified rat cTnI was 50% more reactive than purified human cTnI in the Centaur assay. In the rat, an age- and gender-dependent variation in serum cTnI was found. Male rats aged six and eight months had a 10-fold greater serum cTnI than age-matched females and three-month-old rats. These increases correlated with minimal histopathological change. Isoproterenol-induced serum cTnI increased up to 760-fold the minimal detectable concentration of 0.07 microg/L, within 4-6 h and decreased with a half-life of 6 h, with an expected return to baseline of 60 h. Severity of histopathological change correlated with serum cTnI during the ongoing injury.
Assuntos
Doenças dos Animais/sangue , Animais de Laboratório/sangue , Cardiopatias/veterinária , Medições Luminescentes/veterinária , Troponina I/sangue , Doenças dos Animais/diagnóstico , Animais , Biomarcadores/sangue , Cães , Feminino , Cardiopatias/sangue , Medições Luminescentes/normas , Masculino , Camundongos , Miocárdio/química , Ratos , Sensibilidade e Especificidade , Troponina T/sangueRESUMO
A 2-year-old Sprague-Dawley rat with hindlimb paralysis was diagnosed with a cerebral malignant astrocytoma. The distinctive feature of this astrocytoma was the presence of scattered binucleated cells that contained hypereosinophilic, 1-2 micro m in diameter, cytoplasmic granules. The neoplastic astrocytes stained positively for vimentin (VIM), lysozyme, and phosphotungstic acid hematoxylin (PTAH). Within the binucleated cells, granules stained with PTAH and periodic acid-Schiff (PAS) before and after diastase digestion. Ultrastructurally, neoplastic astrocytes were characterized by cytoplasmic aggregates of electron-dense intermediate filaments consistent with VIM and desmin. The cytoplasm of binucleated cells contained numerous phagolysosomes enlarged by myelin figures and glycoprotein or glycolipid. Intermediate filaments were not present. This is the first description, in the rat, of a neoplasm with features resembling the human granular cell astrocytoma. Our findings suggest that an astrocytic origin should be considered for the binucleated cells in this neoplasm.
Assuntos
Astrocitoma/veterinária , Neoplasias Encefálicas/veterinária , Tumor de Células Granulares/veterinária , Doenças dos Roedores/patologia , Animais , Astrocitoma/patologia , Neoplasias Encefálicas/patologia , Tumor de Células Granulares/patologia , Imuno-Histoquímica , Microscopia Eletrônica , Muramidase , Ácido Fosfotúngstico , Ratos , Ratos Sprague-Dawley , Telencéfalo/ultraestrutura , VimentinaRESUMO
Euthymic BALB/c and athymic nude BALB/c mice aged 3-8 days were infected intraperitoneally with Mycobacterium avium subspecies paratuberculosis (ATCC strain 19698). After euthanasia at 5 months post-inoculation, hepatic granulomas were evaluated by morphometric analysis of digital images captured from light microscopy sections, by electron microscopy and by immunohistochemical methods. Euthymic mice differed from athymic mice in that (1) their hepatic granulomas were smaller, contained fewer bacteria, and produced more inducible nitric oxide synthase, and (2) their hepatic macrophages contained fewer bacteria, a higher percentage of degraded bacteria, and increased numbers of primary lysosomes. The study showed that macrophage activation was markedly less in the T cell-deficient athymic mice than in the euthymic mice.
Assuntos
Mycobacterium avium subsp. paratuberculosis/fisiologia , Óxido Nítrico Sintase/biossíntese , Paratuberculose/enzimologia , Linfócitos T/imunologia , Animais , Animais Recém-Nascidos , Modelos Animais de Doenças , Técnica Indireta de Fluorescência para Anticorpo , Granuloma/enzimologia , Granuloma/parasitologia , Granuloma/patologia , Processamento de Imagem Assistida por Computador , Técnicas Imunoenzimáticas , Fígado/enzimologia , Fígado/imunologia , Fígado/ultraestrutura , Lisossomos/microbiologia , Lisossomos/ultraestrutura , Macrófagos/enzimologia , Macrófagos/imunologia , Macrófagos/ultraestrutura , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Óxido Nítrico Sintase/imunologia , Óxido Nítrico Sintase Tipo II , Paratuberculose/imunologia , Paratuberculose/patologiaAssuntos
Doenças dos Bovinos/diagnóstico , Mycobacterium avium subsp. paratuberculosis/imunologia , Paratuberculose/diagnóstico , Animais , Anticorpos Antibacterianos/análise , Bovinos , Doenças dos Bovinos/imunologia , Formaldeído , Imuno-Histoquímica , Inclusão em Parafina , Paratuberculose/imunologia , Sensibilidade e Especificidade , Preservação de TecidoRESUMO
A 7-month-old foal was admitted to the hospital with a history of lethargy, weight loss, mild diarrhea, and anorexia. A diagnosis of proliferative enteritis caused by Lawsonia intracellularis-like organisms was made after necropsy and histologic examination of the small intestine. Although infection with L intracellularis-like organisms is a rare cause of enteritis in foals, it should be considered in the differential diagnosis, especially if the foal was housed in the proximity of pigs or pig feces. Antemortem diagnosis remains challenging because isolation of the organism in fecal material requires cell culture, and histologic evaluation of intestinal biopsy specimens may be unrewarding because of the lack of information regarding the frequency and distribution of lesions in horses. Alternatively, use of immunochemical stain, dot-blot technique, and polymerase chain reaction provide specific diagnostic tests that can be performed on fecal material. Postmortem diagnosis relies on histologic examination of infected tissues and use of immunofluorescence and polymerase chain reaction.