Heller Laboratory
Stanford University

The Wave – The Golden Cathedral – Coyote Gulch and lots of gravel road travel…

Perfect weather this year.  70s in April, which was a refreshing change from last year’s scorching heat.  Only incident this year, a fall into a cactus 🙁  Well, learning experience this time:  white Elmer type glue and gauze is the proven best method for removing prickly pear cactus spikes (peer reviewed papers exist, well not sure about how serious science this is though…).  Another item to add to the emergency kit…

From Las Vegas, we drove to the Wave (Wire Pass) trailhead and camped a night in the wilderness.  Thanks to Mirko (again!), we won yet another 6-person permit for The Wave. This hike never gets boring.  On the way back, we passed areas that were flooded last time (2015) – just compare the pictures and it becomes clear.  We then drove via Cottonwood Canyon and the Hole in the Rock Road to the trailhead (Egypt trailhead) for the Golden Cathedral.  This was a fun hike through desert, wading through water, and with some areas of lush vegetation.  Just as we settled down at the Golden Cathedral, we observed a group of climbers repelling through the pothole arches from the roof of the cathedral.  The next day, we headed to a long day hike through Coyote Gulch.  This time we did not attempt the dangerous climb out of the canyon, but rather took the long and safe route out via a never ending walk on crunchy gravel through Hurricane Wash.  We all had an excellent time!

Kane Gulch – Grand Gulch – Bullet Canyon … bring water …

It was hot – very hot!  This year we took an early morning flight to Salt Lake City, met up with former lab member Eduardo, who joined us from Boston.  First time with trucks – well, they are useful but not necessarily most comfortable.  After a long drive South, loading of supplies (don’t buy water at a Gas Station ($$$)), we arrived at Kane Gulch Ranger station, the gateway to Grand Gulch.  We deposited the second car “on the other side”, where we planned to hike out of the canyon network via Bullet Canyon. This whole area is called Cedar Mesa. It is quite remote (in fact, we did not see a soul for the whole 3 day backpacking trip).  Did I mention that it was hot 🙂

Grand Gulch has plenty of ancient dwellings that were built and inhabited by ancestral Puebloans from 700 – 2000 years ago.  The whole place is pretty much a huge outdoor museum.  Hiking, by the way, is pretty awesome, except for the occasional bushwhacking through dense “stuff” with really annoying spiky seeds.  Another four learning experiences during this hike a) 1 Gallon of water per day might not be enough in hot weather (just drop some other items, like a hammer! or too much food) – a bottle of wine is of course absolutely necessary.  b) Never pack 1 gallon water canisters from the supermarket.  They tend to develop leaks because they are made from very thin plastic. Spread the water into small packs such as Platypus type bottles (although recently they also seem to be made from cheaper and thinner plastic).  c) IF you have a cell phone with GPS, load a topographical map (set a waypoint at the trailhead(s)) and switch the phone off (no reception anyways) until you need access to the map.  Having map and GPS is very good if the group splits up and you are not sure about where the trailhead with the car is.  And d) Never panic!   Now it is probably obvious that we had some issues on the last day of the hike.  Dehydration is dangerous.  But we all made it back, sound and safe – a little smarter as well.

On the way back to Salt Lake City, we “stopped” at Horseshoe Canyon, hiked to the Great Gallery, and we spend a “Spa” night with a hot shower in Goblin Valley State Park.  All these are beautiful places and well are worth a visit (even repeatedly).

Arches NP

After an early morning flight to Salt Lake City, we loaded up with “survival goods”, stopped at Dead Horse Point SP for a picnic and arrived at Arches NP in the afternoon, checked out Delicate Arch and spent the night at the Devils Garden Campground.  5AM in the morning, we were the first visitors of Devils Garden and did the extended 7+ miles loop.  We loaded up on H2O and food in Moab and then headed South to Canyonlands NP to the Elephant Hill trailhead leading to Chesler Park, our main attraction for 2013!

Chesler Park

Hiking 4 miles mainly uphill into Chesler Park happened in the afternoon and we set up camp in the backcountry.  For the next 2 days, we explored the area, which is quite surreal and beautiful.  On the only full day, we started with the loop trail in the morning and then hiked to Druid Arch in the afternoon (about 17 miles in total).

Goblin Valley and the Great Gallery

Heading back north to Salt Lake City, we stopped at Goblin Valley SP for a quick shower and sight seeing, headed to the backcountry again to camp at the Horseshoe Canyon trailhead for to the Great Gallery.  On the last day, we hiked to the Great Gallery, before heading back to SLC and our flights home.  Around midnight, we touched down, dirty, dusty, tired, but quite happy.

2018 publications:

• Janesick, Amanda S. and Heller, Stefan. Stem Cells and the Bird Cochlea – Where is Everybody?  Cold Spring Harbor Perspectives in Medicine, doi: 10.1101/cshperspect.a033183 (2018). Link to the paper.

In this perspective, we describe different mechanisms of hair cell regeneration in the avian cochlea and argue that there is no doubt that cochlear hair cell regeneration in birds is driven by stem cells.  With all this evidence for stem cells, the question remains:  Where are they?  This short essay pretty much outlines the need for a thorough analysis of hair cell regeneration in the bird inner ear.

• Roccio, Marta; Perny, Michael; Ealy, Megan; Widmer, Hans Ruedi; Heller, Stefan; and Senn, Pascal. Molecular Characterization and Prospective Isolation of Human Fetal Cochlear Hair Cell Progenitors. Nature Communications, 9. Article number: 4027 (2018). Link to the paper at Nature Communications.

This is an important study of the developing human inner ear.  This was not an easy project and I congratulate Marta and Pascal and the rest of the team for making this significant contribution to the field.  The majority of our current understanding how the inner ear develops comes from animal studies and specifically with respect to knowing which genes are involved in human inner ear development, we could only speculate.  This paper validates a lot of existing hypotheses and it also shows that human hair cell progenitors can indeed be identified and coerced to differentiate into cells that one could define as a generic human sensory hair cell.  This does not mean that we are ready to grow such cells for transplantation – it is an early step towards identifying these progenitors cells.  Our lab’s contribution (Megan) was to determine the genes that are expressed in the prospective progenitor cells.  Thank you Pascal and Marta for reaching out and for the successful collaboration.

This paper began with a discussion between Peter and Stefan at the ARO meeting in 2014 where we wondered whether it would be feasible to determine when individual genes become actively expressed during bundle growth and maturation.  Mirko and Rachel were drawn in to identify the best time point during development when hair cells at all stages of development were present in the developing chicken utricle. Mirko profiled >1,000 cells.  Daniel took on the complex challenge of finding an unsupervised computational method to order the single cells along a trajectory that reflects hair bundle maturation and undertook a “tour-de-force” of developing a set of algorithms and their practical realization (CellTrails). The unbiased method consists of innovations and implementations that can be used for a variety of analysis steps including dimensionality reduction, clustering of single cells, determining the number of distinct clusters, as well as the computation and analysis of branching cellular trajectories, including the inference of gene expression dynamics. Moreover, CellTrails has an elegant visualization tool that is scalable and therefore compatible with datasets that contain large cell numbers. Flexible trajectory extraction and alignment of trajectories allowed us to compare the expression dynamics of the development of spatially distinct hair cell types with different hair bundle shapes. Ultimately, our study shows that it is indeed feasible to order developing vestibular hair cells based on the sequential expression of hair bundle genes alone.  Peter’s lab had a impressive toolbox of antibodies to hair bundle proteins that were essential for Mirko’s validation analyses using high resolution microscopy. We utilized the length of the hair bundle as a biological ruler to relate gene expression dynamics with hair bundle length. Our analysis revealed that control of local Ca²⁺ is important for hair bundle growth and moreover, it identified distinct subtypes of vestibular hair cells, which indicates that the number of hair cell subtypes in the utricle is larger than previously thought.

• Scheibinger, M., Ellwanger, D.C., Corrales, C.E., Stone, J.S., and Heller, S. Aminoglycoside damage and hair cell regeneration in the chicken utricle. J Assoc Res Otolaryngology, 19, 17-29. doi: 10.1007/s10162-017-0646-4 (2018). https://link.springer.com/article/10.1007%2Fs10162-017-0646-4

Congratulations to postdoc Mirko for his first first author paper in the Heller laboratory.  This work is the foundation of ongoing work that utilizes single cell transcriptomics to elucidate the mechanisms of hair cell regeneration in the chicken utricle.  It also is the first manuscript of the laboratory on chicken hair cell regeneration, which is currently evolving into a major research focus.  I am grateful to the Hearing Health Foundation and the Stanford Initiative to Cure Hearing Loss for funding this project.  

• Lee, J., Böscke, R., Tang, P.C., Hartman, B.H., Heller, S, and Koehler, K.R. Hair follicle development in mouse pluripotent stem cell-derived skin organoids. Cell Reports, 22, 242-254. doi: 10.1016/j.celrep.2017.12.007. (2018). Link to the paper in Cell Reports

When postdocs Robert Böscke and Byron Hartman showed me that their experiments using the inner ear organoid formation protocol published by Karl Koehler and Eri Hashino a few years ago was producing in relatively robust fashion what looked like mouse hair follicles, I first thought that they are joking.  Well, no joke, the protocol that has now been used by several laboratories to produce inner ear cell types can be tweaked and steered towards production of skin including hair follicles.  Is our lab now in the hair growing business?  Certainly not.  But our collaborators at Indiana University are very serious about pursuing this finding further.  Wouldn’t it be great if this can be translated to humans?  I am happy that we were able to contribute to this story.  What I found fascinating is that the fundamental findings were obtained independently in my laboratory and in Karl’s laboratory – what a great validation!  I am also thankful to all scientists involved in this study for their collaborative spirit.  Instead of competing, we decided to work together.  This saved resources, reduced anxiety, and resulted in my mind in a better story.

2017 publications:

• Meese, S., Cepeda, A.P., Gahlen, F., Adams, C.M., Ficner, R., Ricci, A.J., Heller, S., Reisinger, E., and Herget, M. Activity-dependent phosphorylation by CaMKIIδ alters the Ca2+ affinity of the multi-C2-domain protein otoferlin. Frontiers in Synaptic Neuroscience, 9, 13 (2017). https://www.frontiersin.org/articles/10.3389/fnsyn.2017.00013/full

This work was initiated in the Heller laboratory by postdoctoral fellow Meike Herget, who identified via mass spectrometric analyses proteins that interact with otoferlin.  Initially, this project was a control experiment for the prospective co-immunoprecipitation of proteins that interact with the tip link antigen monoclonal antibody (PCDH15) but as it turned out pursuing the transduction apparatus of hair cells with this strategy was futile.  Meike became the driving force of this project, which led her to collaborate initially with Tony Ricci and later on with Ellen Reisinger’s group.  Congratulations to Sandra Meese for wrapping up the experiments and to Meike and Ellen for their perseverance in getting this story published.

• Chen, C.C., Butz, E.S., Chao, Y.K., Grishchuk, Y., Becker, L., Heller, S., Slaugenhaupt, S.A., Biel, M., Wahl-Schrott, C., and Grimm, C. Small molecules for early endosome-specific patch clamping. Cell Chem Biol, 24, 907-916 (2017). http://www.cell.com/cell-chemical-biology/fulltext/S2451-9456(17)30188-5

Former postdoctoral fellow Christian Grimm continues his quest to elucidate the function of TRPML channels, which are located in endosomes. The lack of refined techniques to assess the function of endosomal ion channels and transporters are one of the existing hurdles in this field. This paper is a fine technical advance describing a strategy for patch-clamp experiments on specific subsets of endosomal vesicles.

2016 publications:

• Ealy, M., Ellwanger, D.C., Kosaric, N., Stapper A.P. & Heller, S. Single-cell analysis delineates a trajectory toward the human early otic lineage. Proceedings of the National Academy of Sciences of the United States of America, 113, 8508-8513 (2016). http://www.pnas.org/content/early/2016/07/08/1605537113

Congratulations to Megan, Daniel, Andres, and Nina.  This manuscript is the product of 3 years of hard work by Megan.  The initial idea was to control the culture conditions of human pluripotent stem cells in a way that minimizes “random” signals among cells.  We hypothesized that monolayer cultures in defined conditions would allow us to guide the stem cells in a somewhat synchronous manner with the ultimate goal of generating early inner ear precursor cells.  We found that this idea, in theory, works well for the early stages of generating the type of tissue that eventually gives rise to the early stages of the inner ear.  What we also realized is, however, that the complex 3-dimensional development of the inner ear as it occurs in the embryo might require a corresponding 3-dimensional environment in the culture dish.  Our story depicts the existing limitations and difficulties for guiding human pluripotent stem cells towards the inner ear lineage.  I believe that this could contribute to the development of novel strategies for generating human inner ear cells in vitro.  It also illustrates that it is a very difficult problem to control the development of a complex cell lineage and highlights the need for novel solutions for this problem.  I am thankful to the editor and the reviewers that they accepted our interpretation of the results and allowed us to publish this “open ended” outcome of a research project in the PNAS.  Innovative data analysis and the idea to compare otic cell phenotypes between human and mouse most certainly helped us to make the manuscript appealing to a more general audience.

• DeJonge, R.E., Liu, X.P., Deig, C.R., Heller, S., Koehler, K.R., and Hashino, E. Modulation of Wnt signaling enhances inner ear organoid development in 3D culture. PLoS One, 11: e0162508 (2016). http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0162508

Generation of sensory hair cells from mouse embryonic and induced pluripotent stem cells is a tricky endeavor.  The cells want to become lots of other things and keeping them on track has been a challenge for many researchers.  In this research led by Karl Koehler and Eri Hashino at Indiana University, and conducted by Rachel DeJonge and other members of the Koehler and Hashino labs, it is shown that addition of a small chemical compound (a Wnt signaling agonist) at a specific time point of the cell guidance protocol substantially improves the yield of inner ear cells.  Our laboratory has made similar observations and we contributed tools (a cell line) and some intellectual input to the story.  Eri and Karl have really shown in recent years that it is possible to reliably generate inner ear cells, including functional hair cells, from embryonic stem cells.  I am happy that our lab was inspiring for this by laying the groundwork specifically with our papers published in 2003 and 2010.

• Cao, J.M., Cheng, X.N., Li, S.Q., Heller, S., Xu Z.G., She, D.L.  Identification of novel MYO18A interaction partners required for myoblast adhesion and muscle integrity. Scientific Reports, 6: 36739 (2016). http://www.nature.com/articles/srep36768

Almost 10 years ago, Heller lab postdoc Zhigang Xu, one of the senior authors of this paper and now leading his own lab at the School of Life Sciences at Shandong University, started to search for protein complexes in hair cell stereocilia. For this, he extensively screened inner ear yeast 2-hybrid libraries and he identified several interesting genes and potentially interacting proteins that might play a role in hair cell function.  Myosin18A was one of the proteins that interested him. We never thought that this initial project and the protein interaction partners that he identified during his postdoc at Stanford would lead him to muscle development in zebrafish. Congratulations to Zhigang and his collaborators to this very meticulously done research.

2015 publications:

• Waldhaus, J.,Durruthy-Durruthy, R. & Heller, S. Quantitative High-Resolution Cellular Map of the Organ of Corti. Cell Reports 11:9 June (2015). http://www.cell.com/cell-reports/abstract/S2211-1247(15)00494-5

It is my opinion that this paper has the potential for a landmark paper in our field and beyond.  First, it clearly shows that single cell gene expression analysis and bioinformatic reconstruction methods are very precise and robust, therefore providing a credible tool for functional and mechanistic analyses.  We will definitely use this technology in coming years to explore cellular and molecular mechanisms that regulate development and possibly also regeneration.  Second, the data shown in this paper clearly demonstrate that conventional in situ hybridizations are a method of the past – good bye in situs!  We initially submitted the manuscript to Neuron where it received one highly constructive review and two reviews indicating that the reviewers were not convinced of the robustness of our methods.  Neuron suggested to move the manuscript to their open-access partner journal Cell Reports.  There, the editor asked us to respond to the critiques raised in the Neuron reviews and the revised paper was accepted.  Beside the high publication cost (5K!), Cell Reports seems to be a reasonable journal – proofing and dealing with the production crew was a breeze, probably also because I was out of town and Jörg and Robert took care of dealing with this process ;-).  Congratulations to both for this fine piece of work!

• Maass, J.C., Gu, R., Basch, M.L., Waldhaus, J., Martin Lopez, E., Xia, A., Oghalai, J.S., Heller, S. & Groves, A.K. Changes in the Regulation of the Notch Signaling Pathway are Temporally Correlated with Regenerative Failure in the Mouse Cochlea. Frontiers in Cellular Neuroscience 9:110 (2015). http://journal.frontiersin.org/article/10.3389/fncel.2015.00110/abstract

Our lab contributed to this paper quantitative single cell measurements of Notch pathway gene expression in the young (P2) and juvenile (P21) mouse cochlea.  What we found supports the main conclusion of the paper that the previously reported loss of responsiveness to inhibition of the Notch pathway in mice is likely explainable (at least in part) by down-regulation of Notch receptors, ligands and effectors.  It does not exclude the Notch pathway as a therapeutic target for hair cell regeneration on mammals, but it suggests that other pathways might have to be targeted either simultaneously or before Notch inhibition.  

• Hartman, B.H., Durruthy-Durruthy, R., Laske, R.D., Losorelli, S. & Heller, S. Identification and Characterization of Mouse Otic Sensory Lineage Genes. Frontiers in Cellular Neuroscience 9:79 (2015). http://journal.frontiersin.org/article/10.3389/fncel.2015.00079/abstract

Byron discovered that much more specific genes exist that can be used as family exclusive markers of the optic lineage than previously thought.  He used existing data from our lab published a few years ago and data generated by Roman Laske, a visiting M.D. from Switzerland and then went on to find that the Fbxo2 gene is specific and highly expressed in cells of the extended pro-sensory region of the cochlear floor that ultimately develop into the organ of Corti and into closely adjacent structures.  The paper is our laboratory’s contribution to a series of inner ear papers in the Frontiers Journal.  Thank you editors and reviewers for inviting us and for constructive reviews.

• Durruthy-Durruthy, R. & Heller, S. Applications for Single Cell Trajectory Analysis in Inner Ear Development and Regeneration. Cell and Tissue Research 361, 49-57 (2015). http://www.ncbi.nlm.nih.gov/pubmed/25532874

This is an invited review that is not really extensive but it shows some of the limitations and directions of the emerging field of temporal transcriptomics.  The development of novel and better bioinformatics analysis methods to order cells as they change their phenotype over time is going to be key to understand regenerative processes in the inner ear.  There is lots of work to be done in this area in future years.

• Durruthy-Durruthy, R., Gottlieb, A. & Heller, S. 3D computational reconstruction of tissues with hollow spherical morphologies using single-cell gene expression data. Nature Protocols 10, 459-474 (2015). http://www.nature.com/nprot/journal/v10/n3/full/nprot.2015.022.html

This paper is a follow up on last year’s paper published in Cell.  We were not able to provide detailed step-by-step instructions about the 3D reconstruction method in the original publication, but we thought that the method would be useful to others.  3 of the 4 reviewers agreed with this view, but it took a while to convince the fourth reviewer.  If one follows the topics of the lab’s publications, there is an emerging theme of combining novel ideas for bioinformatics data analysis and visualization with important questions in inner ear biology.  The current revolution of single cell transcriptomics is going to impact our field enormously.

2014 publications:

• Ronaghi, M., Nasr, M., Ealy, M., Durruthy-Durruthy, R., Waldhaus, J., Diaz, G.H., Joubert, L.M., Oshima, K. & Heller, S. Inner Ear Hair Cell-Like Cells from Human Embryonic Stem Cells. Stem Cells and Development (2014). http://www.ncbi.nlm.nih.gov/pubmed/24512547

In this first publication of 2014, we show that it is generally possible to guide human embryonic stem cells toward the inner ear lineage and to generate otic progenitor cells that can differentiate into hair cell-like cells. We also uncovered many limitations of the technology that need to be addressed in future research. Overall, this paper is the product of many years of hard research; quite a difficult project, when compared to other fields such as generation of neurons or muscle cells. History of the paper and review process: We started with submitting the manuscript to PNAS and Stem Cells. Both journals did not review the manuscript, perhaps because we were very clear that the technology has certain limitations. Rewriting parts of the Abstract and Discussion as well as pointing out to the editor of Stem Cells & Development that this paper is the first one to describe the generation of human inner ear sensory hair cells from embryonic stem cells helped. Two reviewers were very positive and the manuscript was accepted after minor revisions.

• Aguilar, A., Becker, L., Tedeschi, T., Heller, S., Iomini, C. & Nachury, M.V. Alpha-Tubulin K40 Acetylation is Required for Contact Inhibition of Proliferation and Cell-Substrate Adhesion. Molecular Biology of the Cell (2014). http://www.ncbi.nlm.nih.gov/pubmed/24743598

• Slattery, E.L., Oshima, K., Heller, S. & Warchol, M.E. Cisplatin Exposure Damages Resident Stem Cells of the Mammalian Inner Ear. Developmental Dynamics (2014). http://www.ncbi.nlm.nih.gov/pubmed/24888499

• Durruthy-Durruthy, R., Gottlieb, A., Hartman, B.H., Waldhaus, J., Laske, R.D., Altman, R. & Heller, S. Reconstruction of the Mouse Otocyst and Early Neuroblast Lineage at Single Cell Resolution. Cell 157, 964-978 (2014). http://http://www.cell.com/cell/abstract/S0092-8674(14)00411-5

We started this project about 1½ years ago and as outlined in the blog post of Apr 24, this paper marks a new focus and direction of research in our laboratory. This focus builds on single cell analysis technology to address important questions in development and regeneration. The history of this manuscript is short (thank god!): The manuscript went directly to Cell in early Dec 2013 and we received the reviews in early Feb 2014. Generally, one would not be happy when it takes 8 weeks to review a manuscript, but there were the holidays and we knew that we had submitted a very long manuscript with plenty of supplementary information, so it was understandable that it took a while. We were happy when we received the reviews: the editor and reviewers really liked the story and we had only to address a few minor critiques plus the task of shorting the text by about 25% so that it fits with the space requirements of the journal. We would like to thank the anonymous reviewers for being excited and supportive of this work. For the laboratory, this paper is surely something special.

A preview putting this publication into a bigger picture perspective was published by Wen, L. and Tang, F. Reconstructing Complex Tissues from Single-Cell Analyses. Cell 157, 771-773 (2014). http://http://www.cell.com/cell/abstract/S0092-8674(14)00542-X

2013 publications:

• Herget, M., Scheibinger, M., Guo, Z., Jan, T.A., Adams, C.M., Cheng, A.G. & Heller, S. A simple method for purification of vestibular hair cells and non-sensory cells, and application for proteomic analysis. PloS One 8, e66026 (2013). http://www.ncbi.nlm.nih.gov/pubmed/23750277

• Volkenstein, S., Oshima, K., Sinkkonen, S.T., Corrales, C.E., Most, S.P., Chai, R., Jan, T.A., van Amerongen, R., Cheng, A.G. & Heller, S. Transient, afferent input-dependent, postnatal niche for neural progenitor cells in the cochlear nucleus. Proceedings of the National Academy of Sciences of the United States of America 110, 14456-14461 (2013). http://www.ncbi.nlm.nih.gov/pubmed/23940359

• Jan, T.A., Chai, R., Sayyid, Z.N., van Amerongen, R., Xia, A., Wang, T., Sinkkonen, S.T., Zeng, Y.A., Levin, J.R., Heller, S., Nusse, R. & Cheng, A.G. Tympanic border cells are Wnt-responsive and can act as progenitors for postnatal mouse cochlear cells. Development 140, 1196-1206 (2013). http://www.ncbi.nlm.nih.gov/pubmed/23444352

• Heller, S. Special issue on inner ear development and regeneration. Hearing Research 297, 1-2 (2013). http://www.ncbi.nlm.nih.gov/pubmed/23276729

• Cao, H., Yin, X., Cao, Y., Jin, Y., Wang, S., Kong, Y., Chen, Y., Gao, J., Heller, S. & Xu, Z. FCHSD1 and FCHSD2 are expressed in hair cell stereocilia and cuticular plate and regulate actin polymerization in vitro. PloS One 8, e56516 (2013). http://www.ncbi.nlm.nih.gov/pubmed/23437151

• Guo, Z., Grimm, C., Becker, L., Ricci, A.J. & Heller, S. A novel ion channel formed by interaction of TRPML3 with TRPV5. PloS One 8, e58174 (2013). http://www.ncbi.nlm.nih.gov/pubmed/23469151

2012 publications:

• Bermingham-McDonogh, O., Corwin, J.T., Hauswirth, W.W., Heller, S., Reed, R. & Reh, T.A. Regenerative medicine for the special senses: restoring the inputs. The Journal of Neuroscience 32, 14053-14057 (2012). http://www.ncbi.nlm.nih.gov/pubmed/23055472

• Grimm, C., Jörs, S., Guo, Z., Obukhov, A.G. & Heller, S. Constitutive activity of TRPML2 and TRPML3 channels versus activation by low extracellular sodium and small molecules. The Journal of Biological Chemistry 287, 22701-22708 (2012). http://www.ncbi.nlm.nih.gov/pubmed/22753890

• Ronaghi, M., Nasr, M. & Heller, S. Concise review: Inner ear stem cells–an oxymoron, but why? Stem Cells 30, 69-74 (2012). http://www.ncbi.nlm.nih.gov/pubmed/22102534

• Volkenstein, S., Kirkwood, J.E., Lai, E., Dazert, S., Fuller, G.G. & Heller, S. Oriented collagen as a potential cochlear implant electrode surface coating to achieve directed neurite outgrowth. European Archives of Oto-Rhino-Laryngology : official journal of the European Federation of Oto-Rhino-Laryngological Societies 269, 1111-1116 (2012). http://www.ncbi.nlm.nih.gov/pubmed/21952794

2011 publications:

• Monk, K.R., Oshima, K., Jörs, S., Heller, S. & Talbot, W.S. Gpr126 is essential for peripheral nerve development and myelination in mammals. Development 138, 2673-2680 (2011). http://www.ncbi.nlm.nih.gov/pubmed/21613327

• Sinkkonen, S.T., Chai, R., Jan, T.A., Hartman, B.H., Laske, R.D., Gahlen, F., Sinkkonen, W., Cheng, A.G., Oshima, K. & Heller, S. Intrinsic regenerative potential of murine cochlear supporting cells. Scientific Reports 1, 26 (2011). http://www.ncbi.nlm.nih.gov/pubmed/22355545

• Sinkkonen, S.T., Starlinger, V., Galaiya, D.J., Laske, R.D., Myllykangas, S., Oshima, K. & Heller, S. Serial analysis of gene expression in the chicken otocyst. Journal of the Association for Research in Otolaryngology : JARO 12, 697-710 (2011). http://www.ncbi.nlm.nih.gov/pubmed/21853378

2010 publications:

• Grimm, C., Jörs, S., Saldanha, S.A., Obukhov, A.G., Pan, B., Oshima, K., Cuajungco, M.P., Chase, P., Hodder, P. & Heller, S. Small molecule activators of TRPML3. Chemistry & Biology 17, 135-148 (2010). http://www.ncbi.nlm.nih.gov/pubmed/20189104

• Diensthuber, M. & Heller, S. [Characterization of stem cells derived from the neonatal auditory sensory epithelium]. HNO 58, 1056, 1058, 1060-1056 (2010). http://www.ncbi.nlm.nih.gov/pubmed/20632158

• Jörs, S., Grimm, C., Becker, L. & Heller, S. Genetic inactivation of Trpml3 does not lead to hearing and vestibular impairment in mice. PloS One 5, e14317 (2010). http://www.ncbi.nlm.nih.gov/pubmed/21179200

• Lee, K.P., Nair, A.V., Grimm, C., van Zeeland, F., Heller, S., Bindels, R.J. & Hoenderop, J.G. A helix-breaking mutation in the epithelial Ca(2+) channel TRPV5 leads to reduced Ca(2+)-dependent inactivation. Cell Calcium 48, 275-287 (2010). http://www.ncbi.nlm.nih.gov/pubmed/21035851

• Oshima, K., Shin, K., Diensthuber, M., Peng, A.W., Ricci, A.J. & Heller, S. Mechanosensitive hair cell-like cells from embryonic and induced pluripotent stem cells. Cell 141, 704-716 (2010). http://www.ncbi.nlm.nih.gov/pubmed/20478259

• Oshima, K., Suchert, S., Blevins, N.H. & Heller, S. Curing hearing loss: Patient expectations, health care practitioners, and basic science. Journal of Communication Disorders 43, 311-318 (2010). http://www.ncbi.nlm.nih.gov/pubmed/20434163

• Saldanha, S.A., Grimm, C., Allais, C., Smith, E., Ouizem, S., Mercer, B.A., Roush, W.R., Heller, S. & Hodder, P. Identification of Selective Agonists of the Transient Receptor Potential Channels 3 (TRPML3), in Probe Reports from the NIH Molecular Libraries Program Bethesda (MD); 2010). http://www.ncbi.nlm.nih.gov/pubmed/24260779

• Saldanha, S.A., Grimm, C., Mercer, B.A., Choi, J.Y., Allais, C., Roush, W.R., Heller, S. & Hodder, P. Campaign to Identify Agonists of Transient Receptor Potential Channels 3 and 2 (TRPML3 & TRPML2), in Probe Reports from the NIH Molecular Libraries Program Bethesda (MD); 2010). http://www.ncbi.nlm.nih.gov/pubmed/22049579

• Xu, Z., Oshima, K. & Heller, S. PIST regulates the intracellular trafficking and plasma membrane expression of cadherin 23. BMC Cell Biology 11, 80 (2010). http://www.ncbi.nlm.nih.gov/pubmed/20958966

2009 publications:

• Brigande, J.V. & Heller, S. Quo vadis, hair cell regeneration? Nature Neuroscience 12, 679-685 (2009). http://www.ncbi.nlm.nih.gov/pubmed/19471265

• Diensthuber, M., Oshima, K. & Heller, S. Stem/progenitor cells derived from the cochlear sensory epithelium give rise to spheres with distinct morphologies and features. Journal of the Association for Research in Otolaryngology : JARO 10, 173-190 (2009). http://www.ncbi.nlm.nih.gov/pubmed/19247714

• Grimm, C., Jörs, S. & Heller, S. Life and death of sensory hair cells expressing constitutively active TRPML3. The Journal of Biological Chemistry 284, 13823-13831 (2009). http://www.ncbi.nlm.nih.gov/pubmed/19299509

• Li, H., Liu, H., Corrales, C.E., Risner, J.R., Forrester, J., Holt, J.R., Heller, S. & Edge, A.S. Differentiation of neurons from neural precursors generated in floating spheres from embryonic stem cells. BMC Neuroscience 10, 122 (2009). http://www.ncbi.nlm.nih.gov/pubmed/19778451

• Oshima, K., Senn, P. & Heller, S. Isolation of sphere-forming stem cells from the mouse inner ear. Methods in Molecular Biology 493, 141-162 (2009). http://www.ncbi.nlm.nih.gov/pubmed/18839346

• Peng, A.W., Belyantseva, I.A., Hsu, P.D., Friedman, T.B. & Heller, S. Twinfilin 2 regulates actin filament lengths in cochlear stereocilia. The Journal of Neuroscience 29, 15083-15088 (2009). http://www.ncbi.nlm.nih.gov/pubmed/19955359

• Samie, M.A., Grimm, C., Evans, J.A., Curcio-Morelli, C., Heller, S., Slaugenhaupt, S.A. & Cuajungco, M.P. The tissue-specific expression of TRPML2 (MCOLN-2) gene is influenced by the presence of TRPML1. Pflugers Archiv : European Journal of Physiology 459, 79-91 (2009). http://www.ncbi.nlm.nih.gov/pubmed/19763610

• Xu, Z., Ricci, A.J. & Heller, S. Rethinking how hearing happens. Neuron 62, 305-307 (2009). http://www.ncbi.nlm.nih.gov/pubmed/19447085

2008 publications:

• D’Hoedt, D., Owsianik, G., Prenen, J., Cuajungco, M.P., Grimm, C., Heller, S., Voets, T. & Nilius, B. Stimulus-specific modulation of the cation channel TRPV4 by PACSIN 3. The Journal of Biological Chemistry 283, 6272-6280 (2008). http://www.ncbi.nlm.nih.gov/pubmed/18174177

• Huang, M., Sage, C., Li, H., Xiang, M., Heller, S. & Chen, Z.Y. Diverse expression patterns of LIM-homeodomain transcription factors (LIM-HDs) in mammalian inner ear development. Developmental Dynamics 237, 3305-3312 (2008). http://www.ncbi.nlm.nih.gov/pubmed/18942141

• Mizoguchi, F., Mizuno, A., Hayata, T., Nakashima, K., Heller, S., Ushida, T., Sokabe, M., Miyasaka, N., Suzuki, M., Ezura, Y. & Noda, M. Transient receptor potential vanilloid 4 deficiency suppresses unloading-induced bone loss. Journal of Cellular Physiology 216, 47-53 (2008). http://www.ncbi.nlm.nih.gov/pubmed/18264976

• Senn, P. & Heller, S. [Stem-cell-based approaches for treating inner ear diseases]. HNO 56, 21-26 (2008). http://www.ncbi.nlm.nih.gov/pubmed/18231694

• Xu, Z., Peng, A.W., Oshima, K. & Heller, S. MAGI-1, a candidate stereociliary scaffolding protein, associates with the tip-link component cadherin 23. The Journal of Neuroscience 28, 11269-11276 (2008). http://www.ncbi.nlm.nih.gov/pubmed/18971469

2007 publications:

• Cuajungco, M.P., Grimm, C. & Heller, S. TRP channels as candidates for hearing and balance abnormalities in vertebrates. Biochimica et Biophysica Acta 1772, 1022-1027 (2007). http://www.ncbi.nlm.nih.gov/pubmed/17300924

• Grimm, C., Cuajungco, M.P., van Aken, A.F., Schnee, M., Jörs, S., Kros, C.J., Ricci, A.J. & Heller, S. A helix-breaking mutation in TRPML3 leads to constitutive activity underlying deafness in the varitint-waddler mouse. Proceedings of the National Academy of Sciences of the United States of America 104, 19583-19588 (2007). http://www.ncbi.nlm.nih.gov/pubmed/18048323

• Heller, S. & O’Neil, R.G. Molecular Mechanisms of TRPV4 Gating, in TRP Ion Channel Function in Sensory Transduction and Cellular Signaling Cascades. (eds. W.B. Liedtke & S. Heller) Boca Raton (FL); 2007). http://www.ncbi.nlm.nih.gov/pubmed/21204503

• Jeon, S.J., Oshima, K., Heller, S. & Edge, A.S. Bone marrow mesenchymal stem cells are progenitors in vitro for inner ear hair cells. Molecular and Cellular Neurosciences 34, 59-68 (2007). http://www.ncbi.nlm.nih.gov/pubmed/17113786

• Martinez-Monedero, R., Oshima, K., Heller, S. & Edge, A.S. The potential role of endogenous stem cells in regeneration of the inner ear. Hearing Research 227, 48-52 (2007). http://www.ncbi.nlm.nih.gov/pubmed/17321086

• Oshima, K., Grimm, C.M., Corrales, C.E., Senn, P., Martinez Monedero, R., Geleoc, G.S., Edge, A., Holt, J.R. & Heller, S. Differential distribution of stem cells in the auditory and vestibular organs of the inner ear. Journal of the Association for Research in Otolaryngology : JARO 8, 18-31 (2007). http://www.ncbi.nlm.nih.gov/pubmed/17171473

• Oshima, K., Teo, D.T., Senn, P., Starlinger, V. & Heller, S. LIF promotes neurogenesis and maintains neural precursors in cell populations derived from spiral ganglion stem cells. BMC Developmental Biology 7, 112 (2007). http://www.ncbi.nlm.nih.gov/pubmed/17935626

• Senn, P., Oshima, K., Teo, D., Grimm, C. & Heller, S. Robust postmortem survival of murine vestibular and cochlear stem cells. Journal of the Association for Research in Otolaryngology : JARO 8, 194-204 (2007). http://www.ncbi.nlm.nih.gov/pubmed/17334849

• Wu, L., Gao, X., Brown, R.C., Heller, S. & O’Neil, R.G. Dual role of the TRPV4 channel as a sensor of flow and osmolality in renal epithelial cells. American journal of physiology. Renal Physiology 293, F1699-1713 (2007). http://www.ncbi.nlm.nih.gov/pubmed/17699550

2006 publications:

• Corrales, C.E., Pan, L., Li, H., Liberman, M.C., Heller, S. & Edge, A.S. Engraftment and differentiation of embryonic stem cell-derived neural progenitor cells in the cochlear nerve trunk: growth of processes into the organ of Corti. Journal of Neurobiology 66, 1489-1500 (2006). http://www.ncbi.nlm.nih.gov/pubmed/17013931

• Cuajungco, M.P., Grimm, C., Oshima, K., D’Hoedt, D., Nilius, B., Mensenkamp, A.R., Bindels, R.J., Plomann, M. & Heller, S. PACSINs bind to the TRPV4 cation channel. PACSIN 3 modulates the subcellular localization of TRPV4. The Journal of Biological Chemistry 281, 18753-18762 (2006). http://www.ncbi.nlm.nih.gov/pubmed/16627472

• Martinez-Monedero, R., Corrales, C.E., Cuajungco, M.P., Heller, S. & Edge, A.S. Reinnervation of hair cells by auditory neurons after selective removal of spiral ganglion neurons. Journal of Neurobiology 66, 319-331 (2006). http://www.ncbi.nlm.nih.gov/pubmed/16408287

• Rivolta, M.N., Li, H. & Heller, S. Generation of inner ear cell types from embryonic stem cells. Methods in Molecular Biology 330, 71-92 (2006). http://www.ncbi.nlm.nih.gov/pubmed/16846017

• Zhao, Y., Wang, Y., Wang, Z., Liu, H., Shen, Y., Li, W., Heller, S. & Li, H. Sonic hedgehog promotes mouse inner ear progenitor cell proliferation and hair cell generation in vitro. Neuroreport 17, 121-124 (2006). http://www.ncbi.nlm.nih.gov/pubmed/16407756

2005 publications:

• Li, H., Corrales, C.E., Wang, Z., Zhao, Y., Wang, Y., Liu, H. & Heller, S. BMP4 signaling is involved in the generation of inner ear sensory epithelia. BMC Developmental Biology 5, 16 (2005). http://www.ncbi.nlm.nih.gov/pubmed/16107213

• Mutai, H., Mann, S. & Heller, S. Identification of chicken transmembrane channel-like (TMC) genes: expression analysis in the cochlea. Neuroscience 132, 1115-1122 (2005). http://www.ncbi.nlm.nih.gov/pubmed/15857715

• O’Neil, R.G. & Heller, S. The mechanosensitive nature of TRPV channels. Pflugers Archiv : European Journal of Physiology 451, 193-203 (2005). http://www.ncbi.nlm.nih.gov/pubmed/15909178

• Oshima, K. & Heller, S. Sound from silence. Nature Medicine 11, 249-250 (2005). http://www.ncbi.nlm.nih.gov/pubmed/15746932

2004 publications:

• Keresztes, G., Martemyanov, K.A., Krispel, C.M., Mutai, H., Yoo, P.J., Maison, S.F., Burns, M.E., Arshavsky, V.Y. & Heller, S. Absence of the RGS9.Gbeta5 GTPase-activating complex in photoreceptors of the R9AP knockout mouse. The Journal of Biological Chemistry 279, 1581-1584 (2004). http://www.ncbi.nlm.nih.gov/pubmed/14625292

• Li, H., Corrales, C.E., Edge, A. & Heller, S. Stem cells as therapy for hearing loss. Trends in Molecular Medicine 10, 309-315 (2004). http://www.ncbi.nlm.nih.gov/pubmed/15242678

• Li, H., Liu, H., Balt, S., Mann, S., Corrales, C.E. & Heller, S. Correlation of expression of the actin filament-bundling protein espin with stereociliary bundle formation in the developing inner ear. The Journal of Comparative Neurology 468, 125-134 (2004). http://www.ncbi.nlm.nih.gov/pubmed/14648695

• Li, H., Liu, H., Corrales, C.E., Mutai, H. & Heller, S. Correlation of Pax-2 expression with cell proliferation in the developing chicken inner ear. Journal of Neurobiology 60, 61-70 (2004). http://www.ncbi.nlm.nih.gov/pubmed/15188273

• Li, H., Liu, H., Sage, C., Huang, M., Chen, Z.Y. & Heller, S. Islet-1 expression in the developing chicken inner ear. The Journal of Comparative Neurology 477, 1-10 (2004). http://www.ncbi.nlm.nih.gov/pubmed/15281076

2003 publications:

• Fuhrmann, S., Stark, M.R. & Heller, S. Expression of Frizzled genes in the developing chick eye. Gene Expression Patterns : GEP 3, 659-662 (2003). http://www.ncbi.nlm.nih.gov/pubmed/12972002

• Keresztes, G., Mutai, H. & Heller, S. TMC and EVER genes belong to a larger novel family, the TMC gene family encoding transmembrane proteins. BMC Genomics 4, 24 (2003). http://www.ncbi.nlm.nih.gov/pubmed/12812529

• Keresztes, G., Mutai, H., Hibino, H., Hudspeth, A.J. & Heller, S. Expression patterns of the RGS9-1 anchoring protein R9AP in the chicken and mouse suggest multiple roles in the nervous system. Molecular and Cellular Neurosciences 24, 687-695 (2003). http://www.ncbi.nlm.nih.gov/pubmed/14664818

• Li, H., Liu, H. & Heller, S. Pluripotent stem cells from the adult mouse inner ear. Nature Medicine 9, 1293-1299 (2003). http://www.ncbi.nlm.nih.gov/pubmed/12949502

• Li, H., Roblin, G., Liu, H. & Heller, S. Generation of hair cells by stepwise differentiation of embryonic stem cells. Proceedings of the National Academy of Sciences of the United States of America 100, 13495-13500 (2003). http://www.ncbi.nlm.nih.gov/pubmed/14593207

• Martemyanov, K.A., Lishko, P.V., Calero, N., Keresztes, G., Sokolov, M., Strissel, K.J., Leskov, I.B., Hopp, J.A., Kolesnikov, A.V., Chen, C.K., Lem, J., Heller, S., Burns, M.E. & Arshavsky, V.Y. The DEP domain determines subcellular targeting of the GTPase activating protein RGS9 in vivo. The Journal of Neuroscience 23, 10175-10181 (2003). http://www.ncbi.nlm.nih.gov/pubmed/14614075

• Mutai, H. & Heller, S. Vertebrate and invertebrate TRPV-like mechanoreceptors. Cell Calcium 33, 471-478 (2003). http://www.ncbi.nlm.nih.gov/pubmed/12765692

2002 publications:

• Heller, S. Application of physiological genomics to the study of hearing disorders. The Journal of Physiology 543, 3-12 (2002). http://www.ncbi.nlm.nih.gov/pubmed/12181277

• Heller, S. Molecular screens for inner ear genes. Journal of Neurobiology 53, 265-275 (2002). http://www.ncbi.nlm.nih.gov/pubmed/12382280

• Heller, S., Bell, A.M., Denis, C.S., Choe, Y. & Hudspeth, A.J. Parvalbumin 3 is an abundant Ca2+ buffer in hair cells. Journal of the Association for Research in Otolaryngology : JARO 3, 488-498 (2002). http://www.ncbi.nlm.nih.gov/pubmed/12072915

• Montell, C., Birnbaumer, L., Flockerzi, V., Bindels, R.J., Bruford, E.A., Caterina, M.J., Clapham, D.E., Harteneck, C., Heller, S., Julius, D., Kojima, I., Mori, Y., Penner, R., Prawitt, D., Scharenberg, A.M., Schultz, G., Shimizu, N. & Zhu, M.X. A unified nomenclature for the superfamily of TRP cation channels. Molecular Cell 9, 229-231 (2002). http://www.ncbi.nlm.nih.gov/pubmed/11864597

• Liedtke, W., Choe, Y., Marti-Renom, M.A., Bell, A.M., Denis, C.S., Sali, A., Hudspeth, A.J., Friedman, J.M. & Heller, S. Vanilloid receptor-related osmotically activated channel (VR-OAC), a candidate vertebrate osmoreceptor. Cell 103, 525-535 (2000). http://www.ncbi.nlm.nih.gov/pubmed/11081638

• Robertson, N.G., Heller, S., Lin, J.S., Resendes, B.L., Weremowicz, S., Denis, C.S., Bell, A.M., Hudspeth, A.J. & Morton, C.C. A novel conserved cochlear gene, OTOR: identification, expression analysis, and chromosomal mapping. Genomics 66, 242-248 (2000). http://www.ncbi.nlm.nih.gov/pubmed/10873378

• Fuhrmann, S., Heller, S., Rohrer, H. & Hofmann, H.D. A transient role for ciliary neurotrophic factor in chick photoreceptor development. Journal of Neurobiology 37, 672-683 (1998). http://www.ncbi.nlm.nih.gov/pubmed/9858267

• Fuhrmann, S., Kirsch, M., Heller, S., Rohrer, H. & Hofmann, H.D. Differential regulation of ciliary neurotrophic factor receptor-alpha expression in all major neuronal cell classes during development of the chick retina. The Journal of Comparative Neurology 400, 244-254 (1998). http://www.ncbi.nlm.nih.gov/pubmed/9766402

• Geissen, M., Heller, S., Pennica, D., Ernsberger, U. & Rohrer, H. The specification of sympathetic neurotransmitter phenotype depends on gp130 cytokine receptor signaling. Development 125, 4791-4801 (1998). http://www.ncbi.nlm.nih.gov/pubmed/9806927

• Heller, S. & Hudspeth, A.J. Two deaf mice, two deaf mice. Nature Medicine 4, 560-561 (1998). http://www.ncbi.nlm.nih.gov/pubmed/9585227

• Heller, S., Sheane, C.A., Javed, Z. & Hudspeth, A.J. Molecular markers for cell types of the inner ear and candidate genes for hearing disorders. Proceedings of the National Academy of Sciences of the United States of America 95, 11400-11405 (1998). http://www.ncbi.nlm.nih.gov/pubmed/9736748

• Robertson, N.G., Lu, L., Heller, S., Merchant, S.N., Eavey, R.D., McKenna, M., Nadol, J.B., Jr., Miyamoto, R.T., Linthicum, F.H., Jr., Lubianca Neto, J.F., Hudspeth, A.J., Seidman, C.E., Morton, C.C. & Seidman, J.G. Mutations in a novel cochlear gene cause DFNA9, a human nonsyndromic deafness with vestibular dysfunction. Nature Genetics 20, 299-303 (1998). http://www.ncbi.nlm.nih.gov/pubmed/9806553

• Holst, A., Heller, S., Junghans, D., Geissen, M., Ernsberger, U. & Rohrer, H. Onset of CNTFRalpha expression and signal transduction during neurogenesis in chick sensory dorsal root ganglia. Developmental Biology 191, 1-13 (1997). http://www.ncbi.nlm.nih.gov/pubmed/9356167

• Rosenblatt, K.P., Sun, Z.P., Heller, S. & Hudspeth, A.J. Distribution of Ca2+-activated K+ channel isoforms along the tonotopic gradient of the chicken’s cochlea. Neuron 19, 1061-1075 (1997). http://www.ncbi.nlm.nih.gov/pubmed/9390519

• Heller, S., Ernsberger, U. & Rohrer, H. Extrinsic signals in the developing nervous system: the role of neurokines during neurogenesis. Perspectives on Developmental Neurobiology 4, 19-34 (1996). http://www.ncbi.nlm.nih.gov/pubmed/9169916

• Heller, S., Finn, T.P., Huber, J., Nishi, R., Geissen, M., Püschel, A.W. & Rohrer, H. Analysis of function and expression of the chick GPA receptor (GPAR alpha) suggests multiple roles in neuronal development. Development 121, 2681-2693 (1995). http://www.ncbi.nlm.nih.gov/pubmed/7671828

• Heller, S., Huber, J., Finn, T.P., Nishi, R. & Rohrer, H. GPA and CNTF produce similar effects in sympathetic neurones but differ in receptor binding. Neuroreport 5, 357-360 (1993). http://www.ncbi.nlm.nih.gov/pubmed/8298104

• Heller, S., Bühler, S., Kilz, S. & Mieschendahl, M. Bioluminescence-based detection of genetically engineered microorganisms in nonsterile river water. Microbial Releases : viruses, bacteria, fungi 1, 35-39 (1992). http://www.ncbi.nlm.nih.gov/pubmed/1341987