Daniel J. Lee, M.D., F.A.C.S.
Associate Professor of Otology and Laryngology, Harvard Medical School
Efferent auditory brainstem circuits
Our laboratory has focused on auditory brainstem circuits that control the middle ear muscle reflexes, one of two major descending systems that provide feedback to the auditory periphery. Collaborating with M. Christian Brown, Ph.D., we are characterizing the anatomy and physiology of the middle ear muscle reflexes in a rat model using retrograde labeling studies as well as lesioning experiments of the cochlear nucleus. We also perform transneuronal tracing of these auditory brainstem pathways using pseudorabies virus (PRV), a powerful neurotropic viral tracer. Our group is using PRV to identify synaptically linked neurons in the CNS that are involved in both the medial olivocochlear and middle ear muscle reflex pathways.
Auditory brainstem implants
My clinical research interests in pediatric and adult cochlear implants have extended to work in the central auditory system as principal investigator and director of the Helene and Grant Wilson Auditory Brainstem Implant (ABI) Program, a multidisciplinary research and clinical effort with collaborators at Mass. Eye and Ear and Massachusetts General Hospital. Our goals are to 1) provide ABIs to patients who are deaf and are not candidates for cochlear implants due to injured or absent auditory nerves (patients with Neurofibromatosis Type 2, cochlear ossification / labyrinthitis ossificans, severe cochlear hypoplasia, or traumatic bilateral auditory nerve injury and 2) conduct basic and clinical research on how to improve the performance of ABIs.
Pediatric Auditory Brainstem Implant (ABI) Clinical Trial at MEEI and MGH
Dr. Lee is now recruiting patients for a new study on ABI surgery in infants and children. Many of these candidates are born without a hearing nerve or a cochlea and cannot receive the cochlear implant or they have failed cochlear implant surgery. The program is closely collaborating with Professor Vittorio Colletti from the University of Verona in Verona, Italy. Professor Colletti is a world-renowned expert in pediatric ABI surgery. Additional information on this pediatric FDA-approved clinical trial (NCT01864291) and the study criteria is available at clinicaltrials.gov.
Adult Auditory Brainstem Implant (ABI) Clinical Trial at MEEI
An adult FDA-approved clinical trial (NCT01736267) was approved in November 2012 and is actively recruiting deaf patients for the ABI who do not have NF2 and are not cochlear implant candidates (or are failed cochlear implant candidates). These candidates have scarring of the inner ear or cochlea from meningitis, otosclerosis, or temporal bone fracture, injury to the auditory nerve, or failed cochlear implant surgery for auditory neuropathy / auditory dyssynchrony.
If you are a patient, parent, referring physician or audiologist who is interested in our pediatric or adult ABI clinical trials, please contact us at (617) 573-3130 or email us at firstname.lastname@example.org.
Optical stimulation of the auditory system
Our recent work in the ABI lab has included the first efforts to optically stimulate neurons in the central auditory system using optogenetics. In contrast to electricity, light offers a theoretical advantage as it can be focused and may allow for the selective activation of hundreds of independent acoustic channels.
Light can depolarize unmodified neurons through thermal and physical means, as has been shown by Richter and colleagues using the delivery of radiant infrared energy to the cochlea (Izzo, Richter et al. 2006; Izzo, Walsh et al. 2008). An alternative approach is the use of optogenetics and light-sensitive proteins that, when delivered through viral vector-mediated gene therapy, can make specific cells activatable or silenceable by multiple colors of low-level visible light (Hirase, et al. 2002; Wells, Kao et al. 2005; Boyden, et al. 2005; Han, et al. 2007; Wells, Thomsen et al. 2007; Chow, et al. 2010).
For our central auditory system research, we employ channelrhodopsin-2 (ChR2), which is a light-gated single-component microbial transmembrane ion channel originally identified in the Chlamydomonas reinhardtii. ChR2 opens to allow ions into the interior of the cell upon stimulation by blue light (Boyden, et al. 2005).
Our lab has teamed with Dr. Edward Boyden and colleagues at the Massachusetts Institute of Technology to demonstrate that adeno-associated viral vector mediated delivery of ChR2 is possible in auditory neurons of the cochlear nucleus (CN) (Fig. 1; Acker et al., 2011).
Figure 1. ChR2 gene delivery and expression in mouse cochlear nucleus. Fluorescence microscopy demonstrating ChR2+ neurons in the cochlear nucleus following direct viral injection (AAV2/8-ChR2 conjugated with green fluorescent protein (GFP)) into dorsal cochlear nucleus (CN) and incubation of 2-4 weeks. Merged image (left panel) showing ChR2 expression in CN neurons (dashed line defines boundary between CN and brainstem), neural tissue expressing ChR2 (middle panel) and neurons of brainstem and cochlear CN defined by nuclear (DAPI) staining. From Darrow et al. 2013.
The goals of our lab are to demonstrate that the mouse cochlear nucleus or central nucleus of the inferior colliculus can be 1) photosensitized with ChR2 using an adeno-associated viral (AAV) vector and 2) activated with blue light to generate auditory responses in vivo.
These experiments may provide the basis for an auditory implant based on light.
Acker, L., Aubin-Pouliot, A., Hancock, K.E., Hauswirth, W., Boyden, E., Brown, M.C., Lee, D.J. (2011) “Channelrhodopsin-2 gene transfection of central auditory neurons: Toward an optical prosthesis.” In: Abstr. Assoc. Res. Otolaryngol. Abstr. #484.
Boyden, E. S., Zhang, F., Bamberg, E., Nagel, G. and Deisseroth, K. (2005). "Millisecond- timescale, genetically targeted optical control of neural activity." Nat Neurosci 8(9): 1263- 1268.
Darrow, K.N., Slama, M., Kempfle, J., Boyden, E., Polley, D., Brown, M.C., Lee, D.J. (2013) “Electrical stimulation of the cochlear nucleus: Effects of location and pulse rate on inferior colliculus responses.” Abstr. Assoc. Res. Otolaryngol. In Press.
Han, X. and Boyden, E. S. (2007). "Multiple-color optical activation, silencing, and desynchronization of neural activity, with single-spike temporal resolution." PLoS One 2(3): e299.
Hirase, H., Nikolenko, V., Goldberg, J. H. and Yuste, R. (2002). "Multiphoton stimulation of neurons." J Neurobiol 51(3): 237247.
Izzo, A. D., Richter, C. P., Jansen, E. D. and Walsh, J. T., Jr. (2006). "Laser stimulation of the auditory nerve." Lasers Surg Med 38(8): 745753.
Izzo, A. D., Walsh, J. T., Jr., Ralph, H., Webb, J., Bendett, M., Wells, J. and Richter, C. P. (2008). "Laser stimulation of auditory neurons: effect of shorter pulse duration and penetration depth." Biophys J 94(8): 31593166.
Wells, J., Kao, C., Jansen, E. D., Konrad, P. and MahadevanJansen, A. (2005). "Application of infrared light for in vivo neural stimulation." J Biomed Opt 10(6): 064003.
Wells, J. D., Thomsen, S., Whitaker, P., Jansen, E. D., Kao, C. C., Konrad, P. E. and MahadevanJansen, A. (2007). "Optically mediated nerve stimulation: Identification of injury thresholds." Lasers Surg Med 39(6): 513526.