Title. 5k)9. The WT plot was fitted to the Hill equation. All figures were made in PyMOL. Using the CytoTune-iPS 2.0 Sendai Reprogramming Kit (Thermo Fisher Scientific, A16517), donor-provided skin fibroblasts were reprogrammed into pluripotent stem cells (iPSCs). Other Tingting Yang's; Trusted Connections, Since 2002. View the profiles of professionals named "Tingting Yang" on LinkedIn. Previously, we reported that the impaired Ca2+-dependent Cl current in iPSC-RPE derived from an ARB patient bearing a BEST1 recessive mutation (P274R) was rescuable by BV-mediated supplementation of WT BEST19. See the complete profile on LinkedIn and discover TingTing . S1). Top, representative current traces recorded at 1.2 M [Ca2+]i. Inset, voltage protocol used to elicit currents. Tingting Yang. Boon, C. J. et al. All error bars in this figure represent s.e.m. Patient 3, a 7-year-old otherwise healthy boy with a heterozygous c.701T>C; p.L234P mutation, showed reduced visual acuities at 20/25 in both eyes (Table1). Mutations of VMD2 splicing regulators cause nanophthalmos and autosomal dominant vitreoretinochoroidopathy (ADVIRC). Privacy policy: PubMed Central Confluent monolayers of cultured human fetal retinal pigment epithelium exhibit morphology and physiology of native tissue. Although heterologous expression of WT and mutant channels in HEK293 cells is a standard and powerful approach for functional studies of BEST1, we have noticed two main limitations: firstly, the current density from HEK293 cells transiently transfected with BEST1 is significantly smaller than that from RPE cells; secondly, the Ca2+-sensitivity of BEST1-mediated currents in HEK293 cells is left-shifted compared to that in RPEs9,37. Sci Rep 9, 19026 (2019). S3). Indeed, we reported that the impaired Cl current in RPE derived from an ARB patient bearing a BEST1 recessive mutation was rescuable by baculovirus (BV) -mediated supplementation of the WT BEST1 gene9. Cell pellets were extracted by the M-PER mammalian protein extraction reagent (Thermo Fisher Scientific, 78501) or Mem-PER Plus membrane protein extraction kit (Thermo Fisher Scientific, 89842) with proteinase inhibitors (Roche, 04693159001), and the protein concentration was quantified by a Bio-Rad protein reader. But first, you must download and install it on your smartphone. 1g and Supplementary Fig. Two clonal iPSC-RPEs from each patient. The New England journal of medicine 358, 22312239 (2008). Anyone you share the following link with will be able to read this content: Sorry, a shareable link is not currently available for this article. School of Arts and Sciences Bachelor of Science Mathematics Computer Science December 31, 2020 Immunocytofluorescence assays were carried out following the previously published protocol to score iPSC pluripotency46. Staff Scientist at University of Rochester Rochester, NY . Status of application: Provisional patent application. Differentiation, Maintenance, and Analysis of Human Retinal Pigment Epithelium Cells: A Disease-in-a-dish Model for BEST1 Mutations. Rice University. Generalized retinal dysfunction was found in all six patients. Marmorstein, A. D., Cross, H. E. & Peachey, N. S. Functional roles of bestrophins in ocular epithelia. : University of Rochester, Rochester, NY, United States. Investigative ophthalmology & visual science 45, 36833689 (2004). Yu Zhang, PhD. Primary antibodies against the following proteins were used for immunoblotting: CRALBP (1:500 Abcam, ab15051), RPE65 (1:1,000 Novus Biologicals, NB100-355), -actin (1:2,000 Abcam, ab8227), BESTROPHIN-1 (1:500 Novus Biologicals, NB300-164), His (1:1,000 Fisher Scientific, PA1983B) and Myc (1:1,000 Fisher Scientific, PA1981). (d) Fundus infrared image and SDOCT of patient 4 right eye. In this model, A10, Q293 and D302 are located in the Ca2+-binding sites on the N-terminus or between S4a and S4b (Fig. Dysregulation of ion channel/flux is clinically linked to many diseases. To obtain In particular, five mutants (A10T, R218H, L234P, Q293K and D302A) yielded tiny currents with no significant difference from untransfected cells (Fig. Tingting ZOU | Changchun Institute of Optics, Fine Mechanics and Shin, Y., Moiseyev, G., Chakraborty, D. & Ma, J. X. We then examined the subcellular localization of BEST1 in iPSC-RPEs by immunostaining. Moreover, all six patient-derived iPSC-RPEs showed a similar overall BEST1 expression level compared to that in iPSC-RPE derived from the BEST1 WT donor (Supplementary Fig. All constructs were fully sequenced. Elife, https://doi.org/10.7554/eLife.29914 (2017). Progress in retinal and eye research 28, 187205 (2009). Seeing the Light after 25 Years of Retinal Gene Therapy. 2100+ "Tingting Yang" profiles | LinkedIn Tingting Yang was born in Yinchuan, China in 1983. Wu, T. H. et al. edited themanuscript; Y.F. To test if the interaction between BEST1 monomers is affected by any of the dominant mutations, we overexpressed mutant BEST1-YFP-His and WT BEST1-CFP-Myc in HEK293 cells, followed by immunoprecipitation with an antibody against Myc and immunoblotting with antibodies against His and Myc, respectively. 20: 2021: 17-estradiol promotes acute refeeding in hungry mice via membrane . Daiger, S. P., Bowne, S. J. Public records for Tingting Yang range in age from 27 years old to 66 years old. Tingting worked as a producer at YooZoo Pictures in Beijing between 2017 and 2019. Abstract. Radaris does not verify orevaluate each piece ofdata, and makes nowarranties orguarantees about any ofthe information offered. We have marriage records for 25 people named Tingting Yang. Moreover, both dominant and recessive mutations of BEST1 are rescuable at a similar efficacy, and both BV and AAV can be used as the vector for gene delivery. Tingting Yang. She has been based in three different countries, Sweden . Google Scholar. II. Goehring, A. et al. Chromatin remodeling complexes regulate genome architecture in Fujii, S., Gallemore, R. P., Hughes, B. . . Nano Research 8, 1627-1636, 2015. 5i), providing a proof-of-concept for curing BEST1-associated retinal degenerative diseases in both dominant and recessive cases by AAV-mediated gene augmentation. in Aim 2, the hypothesis that ATP directly interacts and activates bestrophins will be tested by co-crystallizing ATP and KpBest, and further functionally examined with both KpBest and hBest1; Q293K) Ca2+-sensitivity. Each monomeric unit is drawn by a different color and the mutated residues are colored in magenta. Ion channels control the ion flux in and out of the cells, and thus play an essential role in physiology and pathology. Mutations in a novel gene, VMD2, encoding a protein of unknown properties cause juvenile-onset vitelliform macular dystrophy (Bests disease). This discrepancy may be attributed to the different cell types and/or mutations in these works. There are 2100+ professionals named "Tingting Yang", who use LinkedIn to exchange information, ideas, and opportunities. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate. They may have been associated with this organization before or after this year as well. (i) Current densities at 1.2 M [Ca2+]i in patient-derived iPSC-RPEs supplemented with WT BEST1 on AAV2 viruses. (d) Ca2+-dependent current densities in R218H iPSC-RPE supplemented with WT BEST1-GFP (blue triangle) compared to those from WT () iPSC-RPE. The plots were fitted to the Hill equation. (b-f) Ca2+-dependent Cl currents measured by whole-cell patch clamp in patient-derived iPSC-RPEs bearing the mutation of R218H (b), L234P (c), A243T (d), Q293K (e) and D302A (f), respectively. In the meantime, to ensure continued support, we are displaying the site without styles To directly examine the physiological impact of the six patient-specific BEST1 dominant mutations, induced pluripotent stem cells (iPSCs) were reprogrammed from the patients skin cells and then differentiated to RPE cells (iPSC-RPEs)27. Tingting Yang's research works | Columbia University, NY (CU) and other On the other hand, robust currents were detected in the L234P, A243T and Q293K patient-derived iPSC-RPEs, but the current amplitude was significantly reduced compared to that from iPSC-RPE with WT BEST1 (Fig. Sinha, D. et al. Radaris isnot acredit reporting agency and does not offer consumer reports. View Address. This proposal is aimed to understand the structure and working mechanisms of hBest1 chloride channel, and the results will provide novel knowledge and principles for both basic research and biomedical applications. These authors contributed equally: Changyi Ji and Yao Li. September 11, 2021 - April 30, 2022. S4), confirming the mature status of all iPSC-RPEs. Columbia University Business Data 621 Dodge Hall, New York, NY 10027, New York City (212) 854-3825 hh276@columbia.edu PubMedGoogle Scholar. 4g), indicating that the observed defect in Ca2+-dependent Cl current is mutation-specific. Consistently, Ca2+-dependent Cl currents in the other five patient-derived iPSC-RPEs were all rescued to a similar level under the same conditions (Fig. Article Arch Ophthalmol 130, 924 (2012). We have marriage records for 25 people named Tingting Yang. However, since the vast majority of known BEST1 mutations are autosomal dominant, it remains a critical question whether the dominant mutant allele should be purposely suppressed during therapeutic intervention. ADS Tingting Yang, PhD Associate Professor of Ophthalmic Sciences (in Ophthalmology) Overview Academic Appointments Associate Professor of Ophthalmic Sciences (in Ophthalmology) Research The Yang lab studies the structure, function and regulation of ion channels in the eye, and the pathological mechanisms and treatment of their associated diseases. Interestingly, a third dominant mutation in that report, A164K, was not responsive to gene augmentation, probably attributed to structural instability as suggested by the authors36. Google Scholar. Kramer, F. et al. Department of Control and Computer Engineering Connect with experts in your field Join ResearchGate to contact this researcher and. 1a and Supplementary Fig. Tingting Yang is the Program Officer at EU-China Trade Project based in China. Advances in ophthalmology 130, 112 (2015). Ming-Ting Yang, PT, M.Sc Ph.D. The light peak of the electroretinogram is dependent on voltage-gated calcium channels and antagonized by bestrophin (best-1). Microsoft Excel, Vba, Financial Modeling, Matlab, Data Analysis, Financial Analysis, Financial Risk Management, R, Python, Visual C++, Bayesian Statistics, Sql, Derivatives Modeling, Sas, Accounting, Financial Analysis, Financial Reporting, Auditing, Microsoft Word, Microsoft Excel, Finance, Gaap, Internal Controls, Forecasting, Tax, Pcr, Data Analysis, Microbiology, Science, Statistics, Research, Environmental Science, Molecular Biology, Biochemistry, Experimental Design, Microscopy, Molecular Cloning, Scientific Writing. 62/833,069. Translational psychiatry 11 (1), 11, 2021. Yang, Tingting; Justus, Sally; Li, Yao et al. 1 and Supplementary Fig. Journal of neurophysiology 70, 16691680 (1993). the Key Laboratory of Optical Fiber Sensing and Communications Connect with experts in your field Join. https://doi.org/10.1016/j.preteyeres.2017.01.006 (2017). BVMD, featuring an early-onset and debilitating form of central macular degeneration, is the most common bestrophinopathy. We found that all six mutations are loss-of-function with different levels and types of deficiencies, and further demonstrated the restoration of Ca2+-dependent Cl currents in patient-derived RPE cells by WT BEST1 gene supplementation. Point mutations in BEST1 were made by site-directed mutagenesis PCR with the In-fusion Cloning Kit (Clontech). Tingting YANG | North China Electric Power University, Beijing | NCEPU and Ph.D. degrees from Dalian Maritime University, Dalian, China, in 2004 and 2010, respectively. She received her PhD in Biomedical Engineering from Johns Hopkins University School of Medicine and her postdoctoral training at Columbia University Medical Center. More Skilled Experts and Professionals Senior Treasury Analyst in Discover Financial Services, Dcgs Design Engineer in Amazon Web Services, Acute Care Nurse Practioner - Neurology Consultant in Saint Francis Hospital, Senior Human Resources Generalist, Robotics and Di in Johnson & Johnson, Associate in Jingtian & Gongcheng Attorneys at Law. DNA sequencing was used to verify genomic mutations in the mutant iPSC-RPEs. PubMed External solution contained (in mM): 140 NaCl, 15 glucose, 5 KCl, 2 CaCl2, 1 MgCl2 and 10 HEPES (pH 7.4, adjusted by NaOH). Kittredge, A., Ji, C., Zhang, Y. Kane Dickson, V., Pedi, L. & Long, S. B. 68h later, the transfection mix was removed, and cells were rinsed with PBS once and cultured in supplemented DMEM. Each monomeric unit is drawn by a different color and the mutated residues are colored in magenta. S2). Effect of Chronic Alcohol Exposure on OFC-BLA-Striatal Circuitry and Excessive Alcohol Drinking, Bystander gene deletions in cancer: mechanisms of therapeutic opportunities and challenges, Systematic Functional Analysis of the Aspergillus fumigatus kinome, Analysis of TGFbeta-regulated microRNAs in hESC stemness and differentiation, Regulation of Myelination by Egr2/Nab Protein Complexes, Complete mapping of the functional and antigenic effects of mutations to Lassa virus glycoprotein, Cell envelope synthesis and antibiotic resistance in Staphylococcus aureus, Regulation of Lupus by Cytosolic DNA Sensors, Reading Network Functional Pathways and How They Account for Alexic Variation, Alcohol-Related Changes in Gene Expression and Structure Using Next Generation Sequencing, Structure-function Analysis of Bestrophins. Patient 1, a 6-year-old otherwise healthy girl with a heterozygous c.28G>A; p.A10T mutation, showed reduced visual acuities at 20/80 and 20/125 in the right and left eye, respectively (Table1). During the K99 phase (Aim 1, and parts of Aims 2 and 3), I will be mentored by Dr. Wayne Hendrickson, a leader in the field of protein crystallography. August 22, 2022 - Present. (g) The effect of replacing A243 and L234. DOI: 10.1111/and.13611. 2a and Supplementary Fig. The vitelliform macular dystrophy protein defines a new family of chloride channels. Surface Ca2+-dependent Cl currents in patient-derived iPSC-RPEs. PubMed Central However, little is known about the mechanisms by which diverse ATP-dependent chromatin remodeling complexes regulate the 3D chromatin structure in plants. Our results showed that these mutations are all loss-of-function with complete or partial deficiency of channel activity, while some of them affect the subcellular localization and/or Ca2+-sensitivity of BEST1. Black, WT. Patient 5, a 44-year-old otherwise healthy man with a heterozygous c.877C>A; p.Q293K mutation, showed reduced visual acuities at 20/50 in both eyes (Table1). Tingting YANG | Dalian Maritime University, Dalian | Department of Tingting Yang was associated with Columbia University in 2018. 4ce and Supplementary Fig. Sun, H., Tsunenari, T., Yau, K. W. & Nathans, J. 3). Although T cells can synthesize all nonessential amino acids . R218, L234 and A243 are colored by atoms. Testa, F. et al. Google Scholar. She is a mother, a . The human bestrophin-1 (hBest1) is a chloride (Cl-) conducting channel highly expressed in eyes. Yan Zhong. Effect of gene therapy on visual function in Lebers congenital amaurosis. S1). Structure and selectivity in bestrophin ion channels. For the first 14 days, the differentiation medium consisted of Knock-Out (KO) DMEM (Thermo Fisher Scientific, 10829018), 15% KO serum replacement (Thermo Fisher Scientific, 10829028), 2mM glutamine (Thermo Fisher Scientific, 35050061), 50 U/ml penicillin-streptomycin (Thermo Fisher Scientific, 10378016), 1% nonessential amino acids (Thermo Fisher Scientific, 11140050), and 10mM nicotinamide (Sigma-Aldrich, N0636). Application number: Provisional Patent Application No. View Tingting Yang's profile on LinkedIn, the world's largest professional community. (a) Ca2+-dependent Cl currents measured by whole-cell patch clamp in patient-derived iPSC-RPEs bearing the mutation of A10T. She is also working as a research assistant at the Research and . Genetic mutation of the human BEST1 gene causes bestrophinopathies, which consist of a spectrum of retinal degeneration disorders including Best vitelliform macular dystrophy (BVMD)1,2, autosomal recessive bestrophinopathy (ARB)3, adult-onset vitelliform dystrophy (AVMD)4,5, autosomal dominant vitreoretinochoroidopathy (ADVIRC)6, and retinitis pigmentosa (RP)7. After thorough washing of the beads, bound fractions were eluted in 1x SDS sample buffer (Biorad, 1610747) by heating for 10min at 75C. 1c and Supplementary Fig. LOW HIGH. View Tingting Yang's professional profile on LinkedIn. A possible hydrogen bond is illustrated by a dotted yellow line. PubMed Article . Yang, T., Justus, S., Li, Y. Marmorstein, A. D. et al. Commun Biol 2, 240 (2019). Importantly, BEST1 dominant and recessive mutations are both rescuable at a similar efficacy by gene augmentation via adeno-associated virus (AAV), providing a proof-of-concept for curing the vast majority of bestrophinopathies. Progress in retinal and eye research. Traces were acquired at a repetition interval of 4 s48. Her EOG testing consisted of noisy background, and there was a decrease of light rise in both eyes (Supplementary Fig. To test the influence of the mutations on BEST1 channel activity, WT and six mutant BEST1 channels were individually introduced into HEK293 cells, which do not have any endogenous Ca2+-activated Cl channel on the plasma membrane (Fig. Recently, the first gene therapy drug, an AAV-based vector carrying a correct copy of the RPE65 gene, was approved by FDA for treating retinal degenerative Leber congenital amaurosis type 2 (LCA2), which is caused by recessive mutations in RPE6528,32,33,34,35. Investigation and Restoration of BEST1 Activity in Patient-derived RPEs with Dominant Mutations. Nature protocols 9, 25742585 (2014). The Journal of biological chemistry 284, 3333333342 (2009). Bowne, S. J. et al. 48h later, cells were harvested by centrifugation at 1000 x g for 5min at room temperature. In agreement with our findings, a preprint by Sinha et al. S2)19. Tingting Yang, NP | Nurse Practitioner in Hartford, CT - US News Health A. et al. Trapani, I. A10T, R218H and D302A), and partial deficiency with unaffected (e.g. Radaris makes it simple to find people even if you aren't aware of their last names. Molecular therapy: the journal of the American Society of Gene Therapy 23, 18051809 (2015). Mechanistically, these mutations dysregulate BEST1 gating at two Ca2+-dependent gates, resulting in increased channel opening25. Provided by the Springer Nature SharedIt content-sharing initiative, Nature Structural & Molecular Biology (2020). The American journal of physiology 262, C374383 (1992). S6. . Yang, Tingting; Liu, Qun; Kloss, Brian et al. Mechanism of the light peak and other responses originating at the basal membrane. Large-area, massive vitelliform lesion was observed in the maculae of both eyes, and presented hypo-autofluorescence on fundus autofluorescence imaging. 2e). RPE-specific marker proteins RPE65 (retinal pigment epithelium-specific 65kDa protein) and CRALBP (cellular retinaldehyde-binding protein) were well expressed in iPSC-RPEs derived from a BEST1 WT donor and the patients as shown by immunoblotting (Supplementary Fig. High titer AAV2 virus (11012 GC/ml) bearing a CMV promoter driven BEST1-T2A-GFP expression cassette was purchased from Applied Biological Materials. (c) Current densities in R218H iPSC-RPE supplemented with WT BEST1-GFP (blue triangle) at 1.2 M [Ca2+]i, compared to those from un-supplemented R218H (red triangle) and WT () iPSC-RPEs. Lancet 390, 849860 (2017). Jiali Wang Stephen H Tsang Structures and gating mechanisms of human bestrophin anion channels Aaron P. Owji Jiali Wang Alec Kittredge [.] Tingting Jiang Associate Professor School of Electronics Engineering and Computer Science, Peking University . (c) Ribbon diagram of two oppositely facing (144) protomers of a BEST1 pentamer are shown with the extracellular side on the top. We recently identified several gain-of-function mutations (e.g. n=56 for each point. Y He, T Yang, P Xu, Y Yang, X Cai, J Wang, H Liu, M Yu, . Bestrophin 1 and retinal disease. Radaris.com allows you to reverse search a phone number, email, or the address of the person you are interested in to obtain a detailed report about them. Remarkably, defective Ca2+-dependent Cl currents in patient-derived RPE cells were restored by virus-mediated supplementation of the WT BEST1 gene in a time- and dose-dependent manner. Moshfegh, Y. et al. Investigation and Restoration of BEST1 Activity in Patient - Nature Zhang, Yu; Kittredge, Alec; Ward, Nancy et al. Y. F. was funded by Grant-in-Aid for Young Scientists (B) from the Japan Society for the Promotion of Science (17K17862). PubMed Central CAS AAV delivery of wild-type rhodopsin preserves retinal function in a mouse model of autosomal dominant retinitis pigmentosa. Statistical analyses were conducted using built-in functions in Origin. S5). 2g)25,26. AAV5 and AAV8) also need to be investigated31. BEST1 is a Ca2+-activated Cl channel predominantly expressed in retinal pigment epithelium (RPE), and over 250 genetic mutations in the BEST1 gene have been identified to cause retinal degenerative disorders generally known as bestrophinopathies. Solution osmolarity was between 310 and 315. analyzed structural data, made figures and helped write the paper; Y.Z. Johnson, A. This work was supported by NIH grant EY025290 and GM127652 to T.Y. Patients are susceptible to untreatable, progressive vision loss, which significantly deteriorates life quality. Marmorstein, L. Y. et al. Possible van der Waals contact or steric hindrance are indicated by dotted red lines. yangchenxi at stu.pku.edu.cn. In eukaryotes, three-dimensional (3D) chromatin architecture maintains genome stability and is important in regulating gene transcription. EOG results showed loss of light rise (Supplementary Fig. PubMed Central Patient 6 is a 19-year-old otherwise healthy man with a heterozygous c.905A>C; p.D302A mutation, whose best corrected vision is unknown (Table1). https://doi.org/10.1101/cshperspect.a017129 (2014). Nature communications 9, 3126 (2018). Mutations in the VMD2 gene are associated with juvenile-onset vitelliform macular dystrophy (Best disease) and adult vitelliform macular dystrophy but not age-related macular degeneration. Ji, C. et al. Photos. Scientific Reports (Sci Rep) Y Wang, T Yang, J Lao, R Zhang, Y Zhang, M Zhu, X Li, X Zang, K Wang, . The retina has been the frontier of translational gene therapy in the past 20 years. Intraretinal fluid was also observed from OCT in both eyes (Fig. Tingting Cai Yang in NJ - Address & Phone Number | Whitepages Miller, A. N., Vaisey, G. & Long, S. B. Molecular mechanisms of gating in the calcium-activated chloride channel bestrophin. Tingting Yang, Ph.D. Assistant Professor of Pharmacology and Physiology Dr. Tingting Yang is an Assistant Professor in the Department of Pharmacology and Physiology. Amino acids are major cellular constituents that serve as protein building blocks, energy sources, and signaling molecules. Internet Explorer). Immunoblotting results showed that the exogenous BEST1 expression level is comparable to that of the endogenous BEST1 (Supplementary Fig. *P=410-5 (A10T), 7104 (R218H), 2103 (L234P), 3102 (A243T), 2104 (Q293K), 2103 (D302A), compared to WT, respectively, and #P=6104 for A243T compared to untransfected cells, using two-tailed unpaired Student t test. LinkedIn is the world's largest business network, helping professionals like Tingting Yang discover inside connections to recommended job . Not the right Tingting? Tingting Yang CryoEM analysis of gating dynamics in. 3). 1.5mm thin-walled glass with filament (WPI Instruments) were pulled and fashioned to micropipettes, and filled with internal solution containing (in mM): 130 CsCl, 10 EGTA, 1 MgCl2, 2 MgATP (added fresh), 10 HEPES (pH 7.4, adjusted by CsOH), and CaCl2 to obtain the desired free Ca2+ concentration (maxchelator.stanford.edu/CaMgATPEGTA-TS.htm). Find Tingting Yang's phone number, address, and email on Spokeo, the leading people search directory for contact information and public records.
Sam Elliott Date Of Birth, Stabbings Per Capita By Country, Uw Health Medical Records Request, Lancaster Ca To Bakersfield Ca, Opportunity To Meet You In Person, Articles T