Continuous intraputamenal convection-enhanced delivery in adult rhesus macaques.

OBJECT: Assessing the safety and feasibility of chronic delivery of compounds to the brain using convection-enhanced delivery (CED) is important for the further development of this important therapeutic technology. The objective of this study was to follow and model the distribution of a compound delivered by CED into the putamen of rhesus monkeys. METHODS: The authors sequentially implanted catheters into 4 sites spanning the left and right putamen in each of 6 rhesus monkeys. The catheters were connected to implanted pumps, which were programmed to deliver a 5-mM solution of the MRI contrast agent Gd-DTPA at 0.1 µl/minute for 7 days and 0.3 µl/minute for an additional 7 days. The animals were followed for 28 days per implant cycle during which they were periodically examined with MRI. RESULTS: All animals survived the 4 surgeries with no deficits in behavior. Compared with acute infusion, the volume of distribution (Vd) increased 2-fold with 7 days of chronic infusion. Increasing the flow rate 3-fold over the next week increased the Vd an additional 3-fold. Following withdrawal of the compound, the half-life of Gd-DTPA in the brain was estimated as 3.1 days based on first-order pharmacokinetics. Histological assessment of the brain showed minimal tissue damage limited to the insertion site. CONCLUSIONS: These results demonstrate several important features in the development of a chronically implanted pump and catheter system: 1) the ability to place catheters accurately in a predetermined target; 2) the ability to deliver compounds in a chronic fashion to the putamen; and 3) the use of MRI and MR visible tracers to follow the evolution of the infusion volume over time.

Onset Time and Durability of Huntingtin Suppression in Rhesus Putamen After Direct Infusion of Antihuntingtin siRNA.

One possible treatment for Huntington's disease involves direct infusion of a small, interfering RNA (siRNA) designed to reduce huntingtin expression into brain tissue from a chronically implanted programmable pump. Here, we studied the suppression of huntingtin mRNA achievable with short infusion times, and investigated how long suppression may persist after infusion ceases. Rhesus monkeys received 3 days of infusion of Magnevist into the putamen to confirm catheter patency and fluid distribution. After a 1-week washout period, monkeys received radiolabeled siRNA targeting huntingtin. After 1 or 3 days of siRNA delivery, monkeys were either terminated, or their pumps were shut off and they were terminated 10 or 24 days later. Results indicate that the onset of huntingtin mRNA suppression in the rhesus putamen occurs rapidly, achieving a plateau throughout the putamen within 4 days. Conversely, loss of huntingtin suppression progresses slowly, persisting an estimated 27-39 days in the putamen and surrounding white matter. These findings indicate the rapid onset and durability of siRNA-mediated target gene suppression observed in other organs also occurs in the brain, and support the use of episodic delivery of siRNA into the brain for treatment of Huntington's disease and possibly other neurodegenerative diseases.

Simulated characterization of atherosclerotic lesions in the coronary arteries by measurement of bioimpedance.

FEM software was used to determine the feasibility of characterizing various types of atherosclerotic lesions in vivo. This was accomplished by simulating two electrodes as being attached to an angioplasty balloon in the coronary artery. The electrodes on the "balloon" touched and measured the simulated complex impedance of type III, IV, and Va and Vb lesions, as defined by the American Heart Association (AHA). Additionally, the effect of changes in morphology on the complex impedance was determined for type Va and Vb lesions. The simulations showed that the layer closest to the electrodes had the most significant effect on the measured complex impedance. As a consequence of these simulations, it appears plausible that electrodes could be placed in vivo to determine the characteristics and type of a given atherosclerotic lesion.

Widespread suppression of huntingtin with convection-enhanced delivery of siRNA.

Huntington's disease is an autosomal dominant neurodegenerative disease caused by a toxic gain of function mutation in the huntingtin gene (Htt). Silencing of Htt with RNA interference using direct CNS delivery in rodent models of Huntington's disease has been shown to reduce pathology and promote neuronal recovery. A key translational step for this approach is extension to the larger non-human primate brain, achieving sufficient distribution of small interfering RNA targeting Htt (siHtt) and levels of Htt suppression that may have therapeutic benefit. We evaluated the potential for convection enhanced delivery (CED) of siHtt to provide widespread and robust suppression of Htt in nonhuman primates. siHtt was infused continuously for 7 or 28 days into the nonhuman primate putamen to analyze effects of infusion rate and drug concentration on the volume of effective suppression. Distribution of radiolabeled siHtt and Htt suppression were quantified by autoradiography and PCR, respectively, in tissue punches. Histopathology was evaluated and Htt suppression was also visualized in animals treated for 28 days. Seven days of CED led to widespread distribution of siHtt and significant Htt silencing throughout the nonhuman primate striatum in an infusion rate and dose dependent manner. Htt suppression at therapeutic dose levels was well tolerated by the brain. A model developed from these results predicts that continuous CED of siHtt can achieve significant coverage of the striatum of Huntington's disease patients. These findings suggest that this approach may provide an important therapeutic strategy for treating Huntington's disease.

Two-dimensional axisymmetric model for the sensitivity analysis of a chronic drug infusion into the brain.

A 64-run, 2-level partial factorial experimental analysis was conducted on a 2D axisymmetric finite-element model of the convection-enhanced drug delivery to the parenchyma of the brain. The purpose of this ANOVA analysis was to determine the relative importance of eight factors and their interaction in the volume of distribution for the drug. The analysis revealed that the infusion flowrate and concentration, the overall half-life of the drug and the in vivo effective concentration played an overwhelmingly dominant role in the drug distribution. The results of this analysis will guide the design of an appropriate drug delivery device by focusing research resources on the determined factors of most importance.

Four-point electrode measurement of impedance in the vicinity of bovine aorta for quasi-static frequencies.

Results are presented here of experimental measurements using a four-point electrode technique to measure the complex impedance of bovine aorta submerged in Ringer's solution. Impedance measurements were taken at 250 microm intervals, ranging from 0 (the electrode directly on the surface of the tissue) to 10 mm. Frequencies ranged from 1 kHz to 10 MHz. Throughout this range, the measured impedance changed by an average of 400% when the electrode was 10 mm from the tissue as compared to when the electrode was in direct contact with the tissue. The change in impedance made it possible to determine when the electrode made contact with the arterial wall.