Annual Laboratory Research Progress Report

Research Overview

Stanford OHNS is dedicated to identifying and resolving pathophysiologies associated with the clinical specialty. We have a major focus on inner ear pathologies, but also have strong research across all divisions focused upon identifying and creating the best future treatments. Throughout our department, a multipronged approach is used where clinicians work on a daily basis with primary patients to provide the best service and state of the art care available. Basic scientists work tirelessly in the lab to identify the underlying molecular mechanisms associated with normal and pathological states as a means of developing more focused plans for treatment. Clinician scientists bridge the gap between the bedside and the bench to ensure targeted approaches are being used to identify both the short- and long-term solutions to these pathologies. All of us are dedicated to training and mentoring the next generation of clinicians and scientists who will take the mission of identifying new cures and treatments to greater heights.

This past year has again been quite successful. We have spent time assimilating our new clinician scientists into their positions. We have also hired two new basic scientists, Teresa Nicholson, a senior auditory geneticist with an outstanding track record of identifying critical genes associated with major forms of hearing loss that are functionally critical to sensory cell activity. We have also been fortunate to recruit Daibhid O Maoileidigh, a theoretical physicist who has spent the last ten years devoting his computational prowess to investigating the underpinnings of audition at its fundamental levels. Both of these additions are possible only through the philanthropic support we have been fortunate to receive, and both bring a depth of knowledge and skill in their respective fields that will elevate the entire team to unprecedented heights. The unique collaborative approach within our group is greatly strengthened by these additions and by the philanthropic efforts that make it possible.

2019 brings new excitement as we continue to grow and also to plan for a move into the new Biomedical Innovations building, a state-of-the-art facility that will enhance collaborative efforts. Over the next year we expect to hire at least one more basic science faculty member while also ensuring that our rapidly growing team has the resources and mentoring needed to be successful. Taking advantage of a newly acquired T32 training grant has allowed us to bring highly talented junior clinician scientists into the laboratory, adding yet another dimension to our research team. Through all of these exciting transitions we continue to generate high quality research and identify new targets and sites for intervention. Below is a very brief summary of recent achievements and directions, more details can be found on individual websites as well as the departmental website.

Inner Ear Research

The auditory team includes those whose focus is on the peripheral auditory system: Alan Cheng, Nicolas Grillet, Stefan Heller, Teresa Nicolson, Daibhid O’Maoileidigh and Tony Ricci and middle ear function, Peter Santa Maria.

The overarching goal of the Alan Cheng lab is to protect and regenerate the inner ear and we have continued to make important discoveries in 2018. With the long-term of goal of regenerating the cochlea, we are dissecting the roles of transcription factors and signaling pathways associated with this process. Specifically, we found the surprising interplay between Sox2 and Wnt signaling and also damage in regulating hair cell regeneration and proliferation. This discovery (Atkinson et al., J Clin Inv 2018) allows us to build on the concept that a multipronged molecular approach is promising for stimulating regeneration in the mammalian cochlea.

We continue our efforts in dissecting the roles of Wnt signaling in governing cochlear development. On that end, we are completing projects showing that this pathway is critical in establishing the intricate architecture of the cochlea. In the first project, we found that hair cell orientation (also termed planar cell polarity) involves multiple components, a subset of which requires secreted Wnt proteins for proper function. In parallel, we found that, beta-catenin, a downstream mediator of Wnt signaling regulates the radial patterning of the cochlear organ, an organization critically important for hearing function. Together, these studies provide mechanistic insights into both cochlear development and the potential benefit of manipulating this pathway in regeneration.

In collaboration with Tony Ricci and Hasan DeMirci, we have been using X-ray crystallography to further our understanding of interactions between bacterial ribosomes and the aminoglycoside antibiotics. By comparing conventional and novel aminoglycoside compounds, we found many new conformations representing more complex structure-activity relationships than previously appreciated (O’Sullivan et al., Nucleic Acids 2018).

A third direction in our Lab is to examine molecular mechanisms regulating the regenerating vestibular system in mammals. By studying both the immature and mature regenerating mammalian vestibular system, we found than both systems were able to partially regenerate and recover function. We also found cellular components that are uniquely deficient during regeneration and remarkably different from development. In the coming year, we aim to better define and possibly replace these components in order to augment regeneration.

Mary E. O’Sullivan, Fr´ed´ eric Poitevin, Raymond G. Sierra, Cornelius Gati, E. Han Dao, Yashas Rao, Fulya Aksit, Halilibrahim

Ciftci, Nicholas Corsepius, Robert Greenhouse, Brandon Hayes, Mark S. Hunter, Mengling Liang, Alex McGurk, Paul

Mbgam, Trevor Obrinsky, F´atima Pardo-Avila, Matthew H. Seaberg, Alan G. Cheng, Anthony J. Ricci,* and Hasan DeMirci,* (2018) Aminoglycoside ribosome interactions reveal novel conformational states at ambient temperature. Nucleic Acid Research 1-12

Zahra N. Sayyid_, Grace S. Kim_, and Alan G. Cheng (2018)  Molecular therapy for genetic and degenerative vestibular

 disorders. Co-Otolaryngology Vol 26

Patrick J. Atkinson, Yaodong Dong, Shuping Gu, Wenwen Liu, Elvis Huarcaya Najarro,Tomokatsu Udagawa, and Alan G.

Cheng. (2018) Sox2 haploinsufficiency primes regeneration and Wnt responsiveness in the mouse cochlea. Journal

 of Clinical Investigations.

 

The Nicolas Grillet laboratory uses mouse genetics to investigate inner ear function, and the effects of specific genetic mutations.  The LOXHD1 gene is a novel gene causing hearing loss in humans but with variable manifestation of the phenotype: some patients are affected at birth while others have a later onset and slower progression of hearing loss; all this despite the same nature of the genetic mutation.  Using mouse models, the Grillet group has discovered that regulation of the gene’s specific expression might contribute to this variability.

Furthermore, the Grillet laboratory is characterizing the specific function of the Loxhd1 protein in hair cells, which will provide a major boost to the laboratory in 2018.  Developing novel technologies, like improving scanning electron microscopic imaging of proteins like Loxhd1 combined with simultaneous labeling of the protein with electron-dense particles is allowing unprecedented resolution and insight into the molecular architecture and functioning of this protein. The newly purchased high pressure freezing system (afforded by generous philanthropic money) is enabling the electron microscopic approach to move forward in a way that is beneficial to all members of the department. Also the awarding of an individual investigator NIH award to Nicolas further expands his abilities to push this project forward and attests to his growing preeminence in the auditory field.

Kim J, Xia A, Grillet N, Applegate BE, Oghalai JS. (2018) Osmotic stabilization prevents cochlear synaptopathy after blast trauma.

Proc Natl Acad Sci U S A. May 22;115(21):E4853-E4860. doi: 10.1073/pnas.1720121115. Epub 2018 May 7.

 

2018 was a good year for the Heller laboratory.  Overall, we have very much directed our attention to chicken hair cell regeneration, now with 3 postdocs (Amanda, Nesrine, and Mirko) focusing on this project. Two additional postdocs (Marie and Jiwon) are exploring the generation of chicken:mouse chimeric inner ears.  This exciting new direction is a piece of exploratory research that started in 2018 and if successful, it will allow us to compare the regenerative potential of bird and mammalian cells side-by-side.  In parallel, the Heller lab group has been asking the question how adult mouse cochlear hair cells repair noise-induced damage.  This project is utilizing, like so many in our laboratory, single cell RNA-sequencing and bioinformatics analysis.  Other projects focus on the mechanisms that allow some supporting cells of the newborn mouse to enter a regenerative program, how to guide human embryonic stem cells to adapt an early inner ear phenotype, and how a specific genetic element restricts gene expression in the adult mouse organ of Corti.  We were able to publish our work in good journals in the past year and were able to sustain an appropriate level of funding.

Durruthy-Durruthy R, Sperry ED, Bowen ME, Attardi LD, Heller S, Martin DM. 2018. Single Cell Transcriptomics Reveal Abnormalities in Neurosensory Patterning of the Chd7 Mutant Mouse Ear. Front Genet 9: 473.

Ellwanger DC, Scheibinger M, Dumont RA, Barr-Gillespie PG, Heller S. 2018. Transcriptional Dynamics of Hair-Bundle Morphogenesis Revealed with CellTrails. Cell Rep 23: 2901-2914 e2914.

Hartman BH, Boescke R, Ellwanger DC, Keymeulen S, Scheibinger M, Heller S. 2018. Fbxo2(VHC) mouse and embryonic stem cell reporter lines delineate in vitro-generated inner ear sensory epithelia cells and enable otic lineage selection and Cre-recombination. Dev Biol 443: 64-77.

Janesick AS, Heller S. 2018. Stem Cells and the Bird Cochlea-Where Is Everybody? Cold Spring Harb Perspect Med.

Lee J, Bscke R, Tang PC, Hartman BH, Heller S, Koehler KR. 2018. Hair Follicle Development in Mouse Pluripotent Stem Cell-Derived Skin Organoids. Cell Rep 22: 242-254.

Roccio M, Perny M, Ealy M, Widmer HR, Heller S, Senn P. 2018. Molecular characterization and prospective isolation of human fetal cochlear hair cell progenitors. Nat Commun 9: 4027.

Scheibinger M, Ellwanger DC, Corrales CE, Stone JS, Heller S. 2018. Aminoglycoside Damage and Hair Cell Regeneration in the Chicken Utricle. J Assoc Res Otolaryngol 19: 17-29.

 

The Nicolson Lab has relocated from the Oregon Hearing Research Center to Stanford in the winter of 2019. The prospects for collaborative research at Stanford OHNS are quite high, as the Nicolson lab has substantial overlap in interests with several OHNS labs. Their work focuses on zebrafish animal models of human deafness and vestibular dysfunction. Recent studies have revealed fundamental insights into the assembly of the mechanotransduction complex in sensory hair cells. The Nicolson lab discovered that two factors, one in the secretory pathway (Tomt) and another within the complex (Tmie), are required for targeting mechanosensitive channels to the hair bundle. In both animal models, they found that mature hair cells lacking mechanotransduction over the course of development could be rescued by reintroduction of the wild-type genes. This finding suggests that human patients with progressive forms of hearing loss due to mutations in TOMT or TMIE would be ideal candidates for gene therapy.

 

Dáibhid Ó Maoiléidigh’s laboratory published its first independent study in 2018. This publication described the conditions under which active systems, such as hair cells in the inner ear, can harness environmental fluctuations to better detect weak vibrational input like quiet sounds. The group began several new projects. Work describing how the dynamics of individual hair cells is fundamentally similar to that of the inner ear as a whole was described in an oral presentation at the Association for Research in Otolaryngology’s Midwinter Meeting. At the same meeting, preliminary results were presented from another project in collaboration with Tony Ricci. The goal of this research direction is to understand why mammalian auditory hair bundles, the collection of sensory hairs atop hair cells, differ in structure from vestibular and non-mammalian hair bundles. This work will be submitted for publication in 2019. Another new project with external collaborators is already under review and describes how two active ears can produce sounds through the interaction of both ears. The close agreement between computational modeling and experimental observations in this study provides strong evidence for mechanically active hearing in vertebrates. The Ó Maoiléidigh group has several additional lines of research in cochlear and vestibular mechanics, which will reveal fundamental mechanisms underlying their function.

Ó Maoiléidigh D. (2018) Multiple mechanisms for stochastic resonance are inherent to sinusoidally driven noisy Hopf oscillators, Physical Review E 97:022226

Ó Maoiléidigh D and Hudspeth AJ. (2018) Sinusoidal-signal detection by active, noisy oscillators on the brink of self-oscillation, Physica D: Nonlinear Phenomena 378-379:33

 

The Ricci Lab continues to push forward on a variety of exciting fronts. The major focus remains on hair cell mechanotransduction, i.e. the means by which sound mechanically stimulates the sensory cell and the sensory cell translates this stimulus into an electrical and then chemical signal recognized by the brain. We have two major foci, the first is exploring the role of the lipid bilayer in regulating both the mechanical and biochemical sensitivities of the mechanotransduction process. This novel work is challenging longstanding views of the process and promises to shed new light onto both the normal and pathological condition. This work is led by postdoctoral fellow Shefin George but is quite collaborative and is greatly enhanced by work with Nicolas Grillet, Charles Steele and Dáibhid Ó Maoiléidigh and the long-term collaborations with past postdoctoral fellows Anthony Peng and Thomas Effertz. The second focus is on sensory hair bundle mechanics, where we have shown that unlike every other hair bundle investigated, mammalian cochlear hair bundle stereocilia move more independently. We are trying to understand the functional value and molecular underpinnings of this novel finding. As the properties of the hair bundle are intricately tied to age and noise related damage as well as sensitive to many known genetic disorders, a better understanding of this fundamental process is needed. This work is being led by graduate student Alex Scharr and postdoctoral fellow Yanli Wang but is also greatly enhanced by collaborations with: Anthony Peng, Charles Steele, Sunil Puria and Dáibhid Ó Maoiléidigh.

An offshoot of the mechanotransduction project is the development of nontoxic aminoglycosides. This project has recently made new strides by incorporating X-ray crystallography, and cryoelectron microscopy to directly probe drug binding to its target (led by Hasan DiMirci). The project has recently developed new chemistry for purification of unique drug subtypes (Bob Greenhouse and Nanosyn) that are being tested for ototoxicity (Mary O’Sullivan) and antimicrobial efficacy (Microbiology Institute, Randy Lin and Mary O’Sullivan). Development of a novel tissue culture system has improved throughput on investigating multiple drugs (Mary O’Sullivan) and has led to new insights into drug actions and new modification sites. We are presently starting a new round of synthesis (Nanosyn) that we are confident will need to new drug therapies.

A second major focus of the laboratory is investigating sensory cell afferent fiber communication through ribbon synapses. We have recently published important work on the functional relevance of the ribbon synapse in ELIFE. This was a truly collaborative effort with authors from within our program including Lars Becker, Michael Schnee, Sara Talaei, Mamiko Niwa and also outside of our institution including Bechara Kachar and Mark Rutherford. We are also completing work to investigate potential mechanisms associated with hidden hearing loss providing direct comparisons of afferent fiber properties. Hereto a collaborative effort works best with Mamiko Niwa leading the efforts and Elisabeth Glowatzki and Eric young collaborating. We are also investigating synaptic vesicle trafficking as a potential intervention site for age related and noise induced hearing loss. Mike Schnee and Sara Talaei lead these efforts. In conjunction with these efforts we are developing new drug delivery systems and surgical approaches for these delivery systems. This work also includes new approaches to gene therapy approaches. Sara Talaei and Jinkyung Kim are leading these efforts with support from Mike Schnee, Kyssia Mendoza and Ksenia Aviella Aaron.

Mary E. O’Sullivan, Fr´ed´ eric Poitevin, Raymond G. Sierra, Cornelius Gati, E. Han Dao, Yashas Rao, Fulya Aksit, Halilibrahim

Ciftci, Nicholas Corsepius, Robert Greenhouse, Brandon Hayes, Mark S. Hunter, Mengling Liang, Alex McGurk, Paul

Mbgam, Trevor Obrinsky, F´atima Pardo-Avila, Matthew H. Seaberg, Alan G. Cheng, Anthony J. Ricci,* and Hasan DeMirci,* (2018) Aminoglycoside ribosome interactions reveal novel conformational states at ambient temperature. Nucleic Acid Research 1-12

Nuttall AL, Ricci AJ, Burwood G, Harte JM, Stenfelt S, Cayé-Thomasen P, Ren T, Ramamoorthy S, Zhang Y, Wilson T, Lunner T,

Moore BCJ, Fridberger A. (2018)  A mechanoelectrical mechanism for detection of sound envelopes in the hearing organ. Nature Communication 9: 4175

Morgan CP, Zhao H, LeMasurier M, Xiong W, Pan B, Kazmierczak P, Avenarius MR, Bateschell M, Larisch R, Ricci AJ, Müller U, Barr-

Gillespie PG. TRPV6, TRPM6 and TRPM7 Do Not Contribute to Hair-Cell Mechanotransduction. Front Cell Neurosci. 2018

Feb 20;12:41. doi: 10.3389/fncel.2018.00041. eCollection 2018.

Becker L, Schnee ME, Niwa M, Sun W, Maxeiner S, Talaei S, Kachar B, Rutherford MA, Ricci AJ. (2018)

 The presynaptic ribbon maintains vesicle populations at the hair cell afferent fiber synapse. Elife. 2018 Jan 12;7. pii:

e30241. doi: 10.7554/eLife.30241.

Effertz T, Becker L, Peng AW, Ricci AJ. (2018) Phosphoinositol-4,5 Bisphosphate regulates Audditory Hair-Cell

 Mechanotransduction-Channel Pore Properties and fast Adaptation. J Neurosci. 2017 Nov 29;37(48):11632-11646. doi:

10.1523/JNEUROSCI.1351-17.2017. Epub 2017 Oct 24.

 

The Peter Santa Maria laboratory has discovered that a protein named HB-EGF can initiate keratinocyte proliferation and migration and could be a topical treatment to heal chronic ear drum perforations. This treatment has been licensed to Astellas Pharmaceuticals with a first in human study planned in just over a year. The Santa Maria lab is now focusing on wound healing and immune mechanisms that occur during early Chronic Suppurative Otitis Media, or chronically infected middle ears which are the leading cause of permanent hearing loss in the developing world. Through this study they are studying novel ways to disrupt the cycle of infection including new antimicrobials for use in the ear. The lab also repurposed HB-EGF together with a new mucoadhesive drug delivery system they have developed to prevent post tonsillectomy bleeding and to prevent oral mucositis after chemoradiotherapy.

Beswick DM, Santa Maria C, Ayoub NF, Capasso R, Santa Maria PL. Epithelial separation theory for post-tonsillectomy secondary hemorrhage: evidence in a mouse model and potential heparin-binding epidermal growth factor-like growth factor therapy. Eur Arch Otorhinolaryngol. 2018 Feb;275(2):569-578.

 

The Tulio Valdez laboratory is focused upon the development of new otoscopic technology for diagnosis of middle ear disorders, particularly in pediatric patients.  This includes the use Shortwave infrared imaging for the detection of middle ear effusions and cholesteatoma. Other research explores the use oral films for readily available FDA approved dyes to assess inflammation in the middle ear using Near infrared imaging.  The Valdez group also works on developing and further refining pediatric surgical simulators.  These surgical simulators were featured this year in courses in Israel, Sweden and the United States.

Valdez TA, Carr JA, Kavanagh KR, Schwartz M, Blake D, Bruns O, Bawendi M. Initial findings of shortwave infrared otoscopy in a pediatric population. Int J  Pediatr Otorhinolaryngol. 2018 Nov;114:15-19.

Pandey R, Zhang C, Kang JW, Desai PM, Dasari RR, Barman I, Valdez TA. Differential diagnosis of otitis media with effusion using label-free Raman spectroscopy: A pilot study. J Biophotonics. 2018 Jun;11(6):e201700259. doi: 10.1002/jbio.201700259. Epub 2018 Jan 17.

NIH T32 Auditory Research Program for Residents and Fellows

Grace Kim is a T32 Clinician fellow doing research to develop a working model of regeneration of the mammalian vestibular organ. Grace has been characterizing how neural wiring with sensory cells is re-established. Using fate-mapping approaches, she aims to define the synaptic elements of surviving hair cells and regenerating hair cells as the organ regains function. Her committee includes Alan Cheng, Anthony Ricci, Lloyd Minor and Dáibhid Ó Maoiléidigh. Her work represents how these positions are an excellent means of bringing collaborators from across disciplines together to focus on projects related to inner ear disorders.

Ksenia Aviella Aaron is another clinician fellow whose project focusses upon developing gene therapy approaches to restoring or repairing genetically driven inner ear dysfunction. Unlike other approaches, Ksenia is studying how the properties of the inner ear can influence gene expression while also taking advantage of the strengths in gene therapy at Stanford to identify the best intervention paradigm. She also is investigation the tmprss gene which targets multiple cell types in as yet unidentified manner and so is an extremely difficult disorder to reverse. Success with this gene will open the door to therapeutic interventions across many cell types and time points. Ksenia is being supervised by Anthony Ricci, Alan Cheng, Mark Kay and Nicolas Grillet. Hereto bringing in technical expertise on gene therapy is a major strength of her work.

Taha Jan is a third T32 clinician-scientist fellow who is taking advantage of the fact that the mouse utricle can spontaneously regenerate lost hair cells. He is identifying drivers of mammalian hair cell regeneration by applying single cell RNA sequencing technologies to reveal the transcriptome of hair cell progenitors. Identifying these naturally occurring biochemical pathways, he plans on identifying small molecule activators of these pathways to try and stimulate regeneration in mammalian cochlea. Taha too has a committee of Stefan Heller, Alan Cheng, Roel Nusse and Andrew Huberman. The last two committee members are experts in WNT signaling and retinal tissue regeneration, respectively, but work largely outside of the inner ear. Thus, Taha also serves the important role of liaison between Stanford laboratories both within and across departments.

Jason Qian is the final T32 clinician fellow. His work investigates the role of hearing loss in early onset cognitive function. This project brings a new level of inner ear research to our program by studying how peripheral hearing loss can lead to early onset memory loss, dementia and Alzheimer’s disease. Using recently developed mouse models to create time hearing loss. Jason has yet to create his committee but will take advantage of the strong neuroscience behavioral core to move this project forward and to bring new approaches to inner ear research.

Head and Neck Cancer Research

The John Sunwoo laboratory is focused on understanding how the immune system recognizes and interfaces with head and neck cancer and on ways to translate this to the clinical care of patients. Recently, they have been studying the immune cells present within the tumor microenvironment. They have discovered some tumor cell-intrinsic mechanisms can influence how immune cells are recruited into the tumor. In addition, they have discovered that a special lymphocyte population, called natural killer (NK) cells can be directed by the tumor microenvironment to convert from an anti-tumor phenotype into a pro-tumor phenotype. The understanding of these processes will hopefully lead to novel strategies for therapy. In addition to these studies, the Sunwoo laboratory, along with collaborators at Stanford, has also identified a novel subtype of oral cancers that seem to be unrelated to the standard risk factors, like tobacco use and the human papilloma virus. This subtype seems to affect younger patients and more females than traditional head and neck cancer. The Sunwoo laboratory is studying potential drivers of this subgroup and ways to treat it more specifically.

There have been no improvements in surgical margin rates for forty years and there are currently no methods for intraoperative identification or visualization of pancreatic cancer.  In the Rosenthal Lab we are conducting clinical trials that may enable surgeons to see microscopic fragments of tumors for the first time. We hope this will both improve survival and reduce the removal of normal tissues, dramatically enhancing the effectiveness of surgery as a treatment for cancer.  We have been able to make tumors glow in the operating room by injecting a dye a day or two before surgery.  Early clinical data in head and neck cancer, pancreatic cancer, glioblastoma, and lung cancer shows that this technology will allow doctors to see tumor cells in the clinic (1), on the operating table (2), and even after tumor removal when assessing surgical margins (3).

Improving chemotherapy delivery to tumors

We want to understand why targeted therapies (including immunotherapy) only work in 1 out of 5 patients. We are focused on measuring the successful delivery of antibodies brain and pancreatic cancer tumor.  There has been significant controversy whether large molecules such as antibodies can pass through the dense stroma and poor vascularity in pancreatic cancer or through the blood brain barrier. We propose to test this hypothesis in humans using a novel technique to quantitate antibody delivery to the tumor compared to normal tissues.  We propose to determine biologic factors, microenvironment factors and radiological markers associated with improve delivery.

Miller SE, Tummers WS, Teraphongphom N, van den Berg NS, Hasan A, Ertsey RD, Nagpal S, Recht LD, Plowey ED, Vogel H, Harsh GR, Grant GA, Li GH, Rosenthal EL. First-in-human intraoperative near-infrared fluorescence imaging of glioblastoma using cetuximab-IRDye800.    J Neurooncol. 2018 Apr 6.

Bogyo M, Yim JJ, Rosenthal EL.  New Blood Test SEEKs To Detect and Localize Cancer before It's Too Late.  Biochemistry. 2018 Mar 13;57(10):1561-1562.

Zhang RR, Schroeder AB, Grudzinski JJ, Rosenthal EL, Warram JM, Pinchuk AN, Eliceiri KW, Kuo JS, Weichert JP. Beyond the margins: real-time detection of cancer using targeted fluorophores.  . Nat Rev Clin Oncol. 2017; 14:347-364.

Gao RW, Teraphongphom N, de Boer E, van den Berg NS, Divi V, Kaplan MJ, Oberhelman NJ, Hong SS, Capes E, Colevas AD, Warram JM, Rosenthal EL.  Safety of panitumumab-IRDye800CW and cetuximab-IRDye800CW for fluorescence-guided surgical navigation in head and neck cancers.  Theranostics. 2018; 8(9):2488-2495.

Tummers WS, Miller SE, Teraphongphom NT, Gomez A, Steinberg I, Huland DM, Hong S, Kothapalli SR, Hasan A, Ertsey R, Bonsing BA, Vahrmeijer AL, Swijnenburg RJ, Longacre TA, Fisher GA, Gambhir SS, Poultsides GA, Rosenthal EL.   Intraoperative Pancreatic Cancer Detection using Tumor-Specific Multimodality Molecular Imaging.  Ann Surg Oncol.2018; 139(1): 135–143.

Sinonasal Research

The Nayak Research lab studies Upper Airway Stem Cell Research: CRISPR/CAS9 editing of the CFTR gene locus in upper airway stem cells: Our previous years of research to isolate, define, grow and harness human upper airway basal cells (UABCs) towards the stem-cell based treatment of human disease has recently made exciting translational strides. In mid-2017, we received $2.2M of support from the California Institute of Regenerative Medicine (CIRM) for a collaborative effort (Laboratories of Nayak, Porteus, Kuo and Desai at Stanford) to apply CRISPR gene-editing technology to correct the defective cystic fibrosis (CF) CFTR gene in UABCs to restore chloride transport in these respiratory cells.  Since then, this collaborative team in 2018 has harnessed UABCs to make significant progress to establish: 1) efficient methods to expand the human UABCs in vitro; 2) technologies to edit CFTR in live UABC stem cells; 3) restoration of CFTR function without cell death; 4) methods to re-transplant corrected human UABC stem cells back into living systems. With this foundation, we are now submitting our findings for publication to high-impact factor journals, while seeking extramural grants/funding to advance our trajectory and translational capabilities in this new area of CF treatment.

A second area of interest for the Nayak lab is Upper Airway Immunology Research: Molecular Endotyping of Chronic Rhinosinusitis (CRS) using Multiplexed Ion Beam Imaging (MIBI). CRS is a common disorder that leads to reduced QOL and billions of dollars in costs to patients, yet the factors that drive CRS are poorly understood. Merging our Department’s reputation for advanced care of CRS patients, as well as the greater Stanford research biosphere, we have engineered arrays of sinonasal tissues in order to define the cellular and molecular composition of human upper airway diseases using state-of-the-art MIBI technology. This detailed project, which requires 100’s of tissue samples and elaborate, automated imaging platforms, is now established to the point that extramural applications for funding can be pursued. This work is being advanced in collaboration with the laboratory of Dr. Garry Nolan at Stanford.

 

Laryngeal Research

Voice disorders affect millions of people every year and have a devastating impact on communication and quality of life. Elizabeth DiRenzo’s laboratory is focused on laryngeal mucosal biology. Specifically, we investigate the cellular and molecular events leading to the development of voice disorders and seek to identify unique mechanisms involved in protection of the vocal fold mucosa from injury. Over the past year, we have characterized the multitude of cellular and molecular changes that occur in the laryngeal mucosa of mice and laryngeal cell cultures exposed to conventional cigarette smoke. Electronic (e)-cigarettes, also known as electronic nicotine delivery systems (ENDS), have rapidly become one of the most popular inhalants worldwide. Similar to conventional cigarettes smoke, ENDS-generated vapor is inhaled directly and mucosal surface of the respiratory tract, including the larynx, are the first set of tissues to receive this assault.  Despite the widespread use of ENDS, their effects on laryngeal health is unknown. We have also developed mouse and cell culture models of ENDS exposure that can be used to study the pathophysiological changes that may occur in the larynx of human ENDS users. Early findings demonstrate the ENDS vapor is toxic to laryngeal cells and induces changes to the laryngeal mucosa consistent with inflammatory laryngeal disease. Identifying the effects of ENDS on biological mechanisms that contribute to the development of laryngeal disease is of major significance for making informed clinical recommendations regarding the use of these devices.

Facial Nerve Regeneration Research

Facial paralysis is a debilitating condition that affects approximately 125,000 people in the United States of year. Despite excellent surgical technique, we are unable to restore perfect symmetry to patients with facial paralysis. The mission of Dr. Jon-Paul Pepper’s laboratory is to improve surgical treatment for facial paralysis by identifying strategies that augment facial nerve regeneration after injury. To do this, Dr. Pepper explores the regenerative cues that the body uses to restore tissue after nerve injury. His recent work, featured on the cover of Experimental Neurology, demonstrates a surprising role played by fibroblasts during nerve regeneration after injury. Through the use of cutting edge genetic labeling techniques, Dr. Pepper described the key signaling pathway that activates these cells, and is now manipulating this pathway with small molecules. This pathway, called the Hedgehog signaling pathway, is critical for neurodevelopment and the creation of new blood vessels. It appears that after nerve injury, many of the signals that direct the development of the nervous system are also needed to regenerate it. Importantly, this may provide a means of providing a more robust vascular supply to repaired and/or grafted nerve tissue, which is a significant hurdle in the treatment of all forms of peripheral motor nerve injury. Small molecule treatment for facial paralysis is therefore a promising strategy to improve surgical outcomes. Dr. Pepper’s work will validate the efficacy and safety of this treatment modality in animal models of nerve injury in order to prepare it for clinical use.

Compiled December 19, 2017