Miniature Swine for Preclinical Modeling of Complexities of Human Disease for Translational Scientific Discovery and Accelerated Development of Therapies and Medical Devices.

Noncommunicable diseases, including cardiovascular disease, diabetes, chronic respiratory disease, and cancer, are the leading cause of death in the world. The cost, both monetary and time, of developing therapies to prevent, treat, or manage these diseases has become unsustainable. A contributing factor is inefficient and ineffective preclinical research, in which the animal models utilized do not replicate the complex physiology that influences disease. An ideal preclinical animal model is one that responds similarly to intrinsic and extrinsic influences, providing high translatability and concordance of preclinical findings to humans. The overwhelming genetic, anatomical, physiological, and pathophysiological similarities to humans make miniature swine an ideal model for preclinical studies of human disease. Additionally, recent development of precision gene-editing tools for creation of novel genetic swine models allows the modeling of highly complex pathophysiology and comorbidities. As such, the utilization of swine models in early research allows for the evaluation of novel drug and technology efficacy while encouraging redesign and refinement before committing to clinical testing. This review highlights the appropriateness of the miniature swine for modeling complex physiologic systems, presenting it as a highly translational preclinical platform to validate efficacy and safety of therapies and devices.

Novel anti-Sialyl-Tn monoclonal antibodies and antibody-drug conjugates demonstrate tumor specificity and anti-tumor activity.

Targeted therapeutics that can differentiate between normal and malignant tumor cells represent the ideal standard for the development of a successful anti-cancer strategy. The Sialyl-Thomsen-nouveau antigen (STn or Sialyl-Tn, also known as CD175s) is rarely seen in normal adult tissues, but it is abundantly expressed in many types of human epithelial cancers. We have identified novel antibodies that specifically target with high affinity the STn glycan independent of its carrier protein, affording the potential to recognize a wider array of cancer-specific sialylated proteins. A panel of murine monoclonal anti-STn therapeutic antibodies were generated and their binding specificity and efficacy were characterized in vitro and in in vivo murine cancer models. A subset of these antibodies were conjugated to monomethyl auristatin E (MMAE) to generate antibody-drug conjugates (ADCs). These ADCs demonstrated in vitro efficacy in STn-expressing cell lines and significant tumor growth inhibition in STn-expressing tumor xenograft cancer models with no evidence of overt toxicity.

Historical Incidence of Spontaneous Lesions in Kidneys from Naïve Swine Utilized In Interventional Renal Denervation Studies.

The use of preclinical animal models is integral to the safety assessment, pathogenesis research, and testing of diagnostic technologies and therapeutic interventions. With inherent similarity to human anatomy and physiology, various porcine models have been the preferred preclinical model in some research areas such as medical devices, wound healing, and skin therapies. The porcine model has been the cornerstone for interventional cardiology for the evaluation and development of this catheter-based renal denervation (RDN) therapy. The porcine model provides similar vascular access and renal neurovascular anatomy to humans. In these preclinical studies, the downstream kidneys from treated arteries are assessed for possible histopathological changes in the vessel dependent territories. In assessing renal safety following RDN, it becomes critical to distinguish treatment-related changes from pre-existing background pathologies. The incidence of background pathological changes in porcine kidneys has not been previously established in normal clinically healthy. Samples from the cranial, middle, and caudal portion of 331 naïve kidneys from 181 swine were processed histologically to slides and evaluated microscopically. The most commonly encountered spontaneous changes were chronic pyelonephritis found in nearly half of the evaluated naïve kidneys (∼40 %; score 1 = 91 %, score 2 = 8.4 %, score 3 = 0.76 %) followed by chronic interstitial inflammation in 9.7 % of the kidneys (score 1 = 90.6 %, score 2 = 9.4 %). Interestingly, there were a few rare spontaneous vascular changes that could potentially affect data interpretation in interventional and toxicology studies: arteritis and arteriolar dissection. The presence of pelvic cysts was a common occurrence (6.3 %) in the kidney. The domestic swine is a widely used preclinical species in interventional research, namely in the emerging field of transcatheter renal denervation. This retrospective study presents the historical incidence of spontaneous lesions recorded in the kidneys from naive pigs enrolled in renal denervation studies. There were commonly encountered changes of little pathological consequence such as pyelonephritis or pelvic cysts and rare vascular changes such as arteritis and arteriolar dissection that were of greater potential impact on study data interpretation. These results offer a benchmark by which to gage the potential effect of a procedure or treatment on renal histopathology in swine and assist in data interpretation.

High-throughput, automated extraction of DNA and RNA from clinical samples using TruTip technology on common liquid handling robots.

TruTip is a simple nucleic acid extraction technology whereby a porous, monolithic binding matrix is inserted into a pipette tip. The geometry of the monolith can be adapted for specific pipette tips ranging in volume from 1.0 to 5.0 ml. The large porosity of the monolith enables viscous or complex samples to readily pass through it with minimal fluidic backpressure. Bi-directional flow maximizes residence time between the monolith and sample, and enables large sample volumes to be processed within a single TruTip. The fundamental steps, irrespective of sample volume or TruTip geometry, include cell lysis, nucleic acid binding to the inner pores of the TruTip monolith, washing away unbound sample components and lysis buffers, and eluting purified and concentrated nucleic acids into an appropriate buffer. The attributes and adaptability of TruTip are demonstrated in three automated clinical sample processing protocols using an Eppendorf epMotion 5070, Hamilton STAR and STARplus liquid handling robots, including RNA isolation from nasopharyngeal aspirate, genomic DNA isolation from whole blood, and fetal DNA extraction and enrichment from large volumes of maternal plasma (respectively).

A comparative observational multicenter analysis of the clinical performance of vascular compression devices following transradial arterial access / Análisis comparativo multicéntrico observacional del desempeño clínico de dispositivos de compresión vascular posterior a acceso arterial transradial

Abstract: Introduction: Multiple vascular compression aimed for transradial access have been developed. We aimed to compare the time required to achieve hemostasis in three different radial vascular compression devices. Methods: ST and non-ST elevation MI, unstable and stable angina as well as diagnostic coronary angiograms patients with transradial vascular access (TVA) in 2 centers were enrolled between June 2010-November 2010. Patients were divided according the TVA compression device (TAVCD) used. Group I received TR Band(tm) (Terumo, Tokyo, Japan), Group II received Neptuno(tm) (Biotronik, Berlin, Deutschland) and Group III received Finale(tm) (Merit Medical, South Jordan, UT).Patients were evaluated immediately after TVACD implantation and 24 hour post-procedure follow up. Results: 60 patients were enrolled in this observational study (Group I = 22 patients; Group II = 18; Group III = 20). All patients demonstrated evidence of radial pulse after hemostasis. 28% developed a superficial hematoma (Group I, 31%; Group II, 30%; Group III, 22%). Pain at the access site was uncommon among patients in both immediate and follow-up evaluations, however, 3% (Group I and II) presented paresthesia immediately following the procedure that resolved by the 24-hour evaluation. Conclusion: In our study, all three evaluated radial compression devices successfully achieved hemostasis regardless of the slight alterations of mechanism, yet similarity in aim of non-occlusive compression. Group I (TR band) had a slight increase in compression time recorded but all groups required an approximate three hours to display no evidence of bleeding. None of the patients in the study presented major vascular complications. We consider that further investigation of radial compression devices as compared to manual compression are necessary to evaluate their advantages and may further simplify the procedure

Resumen: Introducción: Múltiples dispositivos de compresión vascular para el acceso radial han sido desarrollados. Nuestro objetivo es comparar el tiempo requerido para lograr la hemostasis con tres diferentes dispositivos de compresión vascular radial. Métodos: Pacientes de angiografía coronaria de diagnóstico, con elevación y no elevación del ST, con angina de pecho inestable y estable, así como pacientes con acceso vascular radial (AVR) en dos centros, fueron incluidos entre junio y noviembre de 2010. Los pacientes fueron divididos según el dispositivo de compresión AVR (DCAVR) usado. Al Grupo I se le dio el TR Band(tm) (Terumo, Tokio, Japón), el Grupo II recibió el Neptuno(tm) (Biotronik, Berlín, Alemania) y el grupo III recibió el Final(tm) (Merit Medica, South Jordan, UT). Los pacientes fueron evaluados inmediatamente después de la implantación del DCAVR y a las 24 horas de postprocedimiento para seguimiento. Resultados: 60 pacientes fueron incluidos en este estudio observacional (Grupo I = 22 pacientes; Grupo II = 18; Grupo III = 20). Todos los pacientes mostraron evidencia de pulso radial después de la hemostasis. 28% desarrolló un hematoma superficial (Grupo I, 31%; Grupo II, 30%; Grupo III, 22%). El dolor en el sitio de acceso fue poco común entre los pacientes de los dos evaluaciones inmediatas y de seguimiento, sin embargo, el 3% (Grupo I y II) presentó parestesias inmediatamente después del procedimiento que se resolvieron para la evaluación de 24 horas. Conclusión: En nuestro estudio, los tres dispositivos de compresión radiales evaluados lograron con éxito la hemostasis sin importar las pequeñas variaciones del mecanismo, dando resultados similares en el objetivo de la compresión no oclusiva. Grupo I (Banda TR) tuvo un ligero aumento en el tiempo de compresión registrado pero todos los grupos requieren un tiempo aproximado de tres horas para mostrar ninguna evidencia de sangrado. Ninguno de los pacientes en el estudio presentó mayores complicaciones vasculares. Consideramos necesario hacer investigación adicional de los dispositivos de compresión radiales, en comparación con la compresión manual para evaluar sus ventajas y poder simplificar aún más el procedimiento.