Murine Drug Testing and Functional Outcome Facility

The preclinical drug testing facility at CNMC is a state-of-the-art facility that is currently extensively used by the academic and industry groups to evaluate efficacy of various therapeutic interventions in muscular dystrophy mouse models. This facility was initially started with a foundation support of $200,000 of seed funding (FED; www.duchennemd.org) in 2006, and subsequently received $600,000 of matching funds from Children’s National Health System.  This provided the funding to equip the facility and the recruitment of Dr. Kanneboyina Nagaraju from Johns Hopkins Medical School to establish and direct the facility. Over the last three years the facility received more than $2 million in grants and contract support (federal, academic labs, foundations, biotechs, and pharmaceuticals) to perform preclinical studies in mouse models of muscular dystrophy. The facility is equipped with computerized multiple grip strength meters (Columbus Instruments, Columbus OH) to assess forelimb and hindlimb strength, Rotarods (Ugo Basile, VA, Italy) to assess motor co-ordination, open-field digiscan behavioral activity chambers (Omnitech Electronics, Columbus, OH) to assess over 20 different activity measurements such as horizontal activity, vertical activity, total distance, movement time and rest time that represent comprehensive animal behavioral activity, treadmills (Columbus Instruments, Columbus, Ohio) to study the effect of  forced exercise on the disease phenotype, and mouse echocardiography (Vevo 770, VisualSonics, Toronto, Canada) to monitor cardiac function in different disease states and response to experimental therapeutics.

In 2008 we have added computerized voluntary wheel running equipment (MiniMitter, with Vital View software) to facilitate studies addressing the effect of exercise on the disease phenotype in mouse models as well as an in vivo optical imaging system (eXplore Optix imaging system; 470nm & 670nm laser/filter sets) to quantitatively measure inflammation and tissue damage in vivo in these mouse models. In addition the facility has developed collaboration with Dr.Paul Wang at Howard University to perform MRI imaging in muscular dystrophy models (full lists of tests in table). The facility routinely performs H&E, special stains (e.g., Sirus red, Trichrome and Immunohistochemistry for selected protein as well as serum creatine kinase assessments for the mice. All the procedures are currently set-up and methods are routinely used in the facility. The facility director, Dr. Nagaraju, has taken the lead and successfully helped to develop SOPs for preclinical phenotyping methods for neuromuscular disorders in full collaboration with Treat-NMD network, and FED (Nagaraju and Willmann, 2009; Spurney et al., 2009).  

Overall the facility currently maintains 8 different mouse models of muscle diseases and has performed over 40 non-GLP preclinical studies in the last three years. The preclinical tests currently being used are listed in Table 1. This facility is currently heavily utilized both by biotech as well as academic investigators for preclinical testing in mouse models.

Inquiries regarding use of the Core should be directed to Kanneboyina Nagaraju DVM PhD.

Table. 1. Preclinical tests currently available at the CNMC drug testing facility

#

Equipment

Measurement

1

Rotarod (Ugo Basile)

Latency to fall

2

Grip strength meter

Fore and hind limb strength

3

Echocardiography

Anatomical and functional measurements of heart

4

GE eXplore Optix

Muscle damage

5

Animal Activity Monitoring System (VersaMax)

25 parameters including total distance, horizontal and vertical activities

6

In vitro muscle function (Aurora scientific, Inc)

Force measurements

7

Open Treadmill for mice (Columbus Instruments)

Exercise

8

MiniMitter-vital view

Voluntary wheel running

9

Serum chemistry  

Serum enzyme levels-CPK

10

Histological (H&E)

Muscle fiber degeneration /regeneration, inflammation and fibrosis

11

IHC

Trichrome, Sirius red

12

Molecular assays

Study relevant proteins

13

Evans Blue Dye

Necrosis

14

MRI through Howard University

Inflammation, Fibrosis and volume