820 results on '"Klein, Ophir D."'
Search Results
2. The developmental basis for scaling of mammalian tooth size
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Christensen, Mona M, Hallikas, Outi, Roy, Rishi Das, Väänänen, Vilma, Stenberg, Otto E, Häkkinen, Teemu J, François, Jean-Christophe, Asher, Robert J, Klein, Ophir D, Holzenberger, Martin, and Jernvall, Jukka
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Archaeology ,Biomedical and Clinical Sciences ,History ,Heritage and Archaeology ,Dental/Oral and Craniofacial Disease ,Rats ,Mice ,Animals ,Proboscidea Mammal ,Molar ,Shrews ,Body Size ,Cognition ,scaling ,size ,shape ,teeth ,IGF - Abstract
When evolution leads to differences in body size, organs generally scale along. A well-known example of the tight relationship between organ and body size is the scaling of mammalian molar teeth. To investigate how teeth scale during development and evolution, we compared molar development from initiation through final size in the mouse and the rat. Whereas the linear dimensions of the rat molars are twice that of the mouse molars, their shapes are largely the same. Here, we focus on the first lower molars that are considered the most reliable dental proxy for size-related patterns due to their low within-species variability. We found that scaling of the molars starts early, and that the rat molar is patterned equally as fast but in a larger size than the mouse molar. Using transcriptomics, we discovered that a known regulator of body size, insulin-like growth factor 1 (Igf1), is more highly expressed in the rat molars compared to the mouse molars. Ex vivo and in vivo mouse models demonstrated that modulation of the IGF pathway reproduces several aspects of the observed scaling process. Furthermore, analysis of IGF1-treated mouse molars and computational modeling indicate that IGF signaling scales teeth by simultaneously enhancing growth and by inhibiting the cusp-patterning program, thereby providing a relatively simple mechanism for scaling teeth during development and evolution. Finally, comparative data from shrews to elephants suggest that this scaling mechanism regulates the minimum tooth size possible, as well as the patterning potential of large teeth.
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- 2023
3. Epithelial zonation along the mouse and human small intestine defines five discrete metabolic domains
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Zwick, Rachel K., Kasparek, Petr, Palikuqi, Brisa, Viragova, Sara, Weichselbaum, Laura, McGinnis, Christopher S., McKinley, Kara L., Rathnayake, Asoka, Vaka, Dedeepya, Nguyen, Vinh, Trentesaux, Coralie, Reyes, Efren, Gupta, Alexander R., Gartner, Zev J., Locksley, Richard M., Gardner, James M., Itzkovitz, Shalev, Boffelli, Dario, and Klein, Ophir D.
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- 2024
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4. Precise modulation of transcription factor levels identifies features underlying dosage sensitivity
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Naqvi, Sahin, Kim, Seungsoo, Hoskens, Hanne, Matthews, Harold S, Spritz, Richard A, Klein, Ophir D, Hallgrímsson, Benedikt, Swigut, Tomek, Claes, Peter, Pritchard, Jonathan K, and Wysocka, Joanna
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Biological Sciences ,Genetics ,Pediatric ,Dental/Oral and Craniofacial Disease ,Biotechnology ,Pediatric Research Initiative ,Humans ,SOX9 Transcription Factor ,Pierre Robin Syndrome ,Gene Expression Regulation ,Regulatory Sequences ,Nucleic Acid ,Phenotype ,Medical and Health Sciences ,Developmental Biology ,Agricultural biotechnology ,Bioinformatics and computational biology - Abstract
Transcriptional regulation exhibits extensive robustness, but human genetics indicates sensitivity to transcription factor (TF) dosage. Reconciling such observations requires quantitative studies of TF dosage effects at trait-relevant ranges, largely lacking so far. TFs play central roles in both normal-range and disease-associated variation in craniofacial morphology; we therefore developed an approach to precisely modulate TF levels in human facial progenitor cells and applied it to SOX9, a TF associated with craniofacial variation and disease (Pierre Robin sequence (PRS)). Most SOX9-dependent regulatory elements (REs) are buffered against small decreases in SOX9 dosage, but REs directly and primarily regulated by SOX9 show heightened sensitivity to SOX9 dosage; these RE responses partially predict gene expression responses. Sensitive REs and genes preferentially affect functional chondrogenesis and PRS-like craniofacial shape variation. We propose that such REs and genes underlie the sensitivity of specific phenotypes to TF dosage, while buffering of other genes leads to robust, nonlinear dosage-to-phenotype relationships.
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- 2023
5. EPIREGULIN creates a developmental niche for spatially organized human intestinal enteroids
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Childs, Charlie J, Holloway, Emily M, Sweet, Caden W, Tsai, Yu-Hwai, Wu, Angeline, Vallie, Abigail, Eiken, Madeline K, Capeling, Meghan M, Zwick, Rachel K, Palikuqi, Brisa, Trentesaux, Coralie, Wu, Joshua H, Pellon-Cardenas, Oscar, Zhang, Charles J, Glass, Ian A, Loebel, Claudia, Yu, Qianhui, Camp, J Gray, Sexton, Jonathan Z, Klein, Ophir D, Verzi, Michael P, and Spence, Jason R
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Biomedical and Clinical Sciences ,Stem Cell Research ,Regenerative Medicine ,Digestive Diseases ,Stem Cell Research - Embryonic - Human ,Biotechnology ,Clinical Research ,Stem Cell Research - Nonembryonic - Human ,Stem Cell Research - Nonembryonic - Non-Human ,1.1 Normal biological development and functioning ,Underpinning research ,Humans ,Epidermal Growth Factor ,Epiregulin ,Intestines ,Intestinal Mucosa ,Cell Differentiation ,Development ,Human stem cells ,Organogenesis ,Stem cells ,Biomedical and clinical sciences ,Health sciences - Abstract
Epithelial organoids derived from intestinal tissue, called enteroids, recapitulate many aspects of the organ in vitro and can be used for biological discovery, personalized medicine, and drug development. Here, we interrogated the cell signaling environment within the developing human intestine to identify niche cues that may be important for epithelial development and homeostasis. We identified an EGF family member, EPIREGULIN (EREG), which is robustly expressed in the developing human crypt. Enteroids generated from the developing human intestine grown in standard culture conditions, which contain EGF, are dominated by stem and progenitor cells and feature little differentiation and no spatial organization. Our results demonstrate that EREG can replace EGF in vitro, and EREG leads to spatially resolved enteroids that feature budded and proliferative crypt domains and a differentiated villus-like central lumen. Multiomic (transcriptome plus epigenome) profiling of native crypts, EGF-grown enteroids, and EREG-grown enteroids showed that EGF enteroids have an altered chromatin landscape that is dependent on EGF concentration, downregulate the master intestinal transcription factor CDX2, and ectopically express stomach genes, a phenomenon that is reversible. This is in contrast to EREG-grown enteroids, which remain intestine like in culture. Thus, EREG creates a homeostatic intestinal niche in vitro, enabling interrogation of stem cell function, cellular differentiation, and disease modeling.
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- 2023
6. Programming multicellular assembly with synthetic cell adhesion molecules
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Stevens, Adam J, Harris, Andrew R, Gerdts, Josiah, Kim, Ki H, Trentesaux, Coralie, Ramirez, Jonathan T, McKeithan, Wesley L, Fattahi, Faranak, Klein, Ophir D, Fletcher, Daniel A, and Lim, Wendell A
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Biochemistry and Cell Biology ,Engineering ,Biological Sciences ,Biomedical Engineering ,Bioengineering ,Underpinning research ,1.1 Normal biological development and functioning ,Generic health relevance ,Cadherins ,Cell Adhesion ,Cell Adhesion Molecules ,Cell Communication ,Integrins ,Synthetic Biology ,Protein Domains ,Binding Sites ,Cell Engineering ,General Science & Technology - Abstract
Cell adhesion molecules are ubiquitous in multicellular organisms, specifying precise cell-cell interactions in processes as diverse as tissue development, immune cell trafficking and the wiring of the nervous system1-4. Here we show that a wide array of synthetic cell adhesion molecules can be generated by combining orthogonal extracellular interactions with intracellular domains from native adhesion molecules, such as cadherins and integrins. The resulting molecules yield customized cell-cell interactions with adhesion properties that are similar to native interactions. The identity of the intracellular domain of the synthetic cell adhesion molecules specifies interface morphology and mechanics, whereas diverse homotypic or heterotypic extracellular interaction domains independently specify the connectivity between cells. This toolkit of orthogonal adhesion molecules enables the rationally programmed assembly of multicellular architectures, as well as systematic remodelling of native tissues. The modularity of synthetic cell adhesion molecules provides fundamental insights into how distinct classes of cell-cell interfaces may have evolved. Overall, these tools offer powerful abilities for cell and tissue engineering and for systematically studying multicellular organization.
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- 2023
7. A DLG1-ARHGAP31-CDC42 axis is essential for the intestinal stem cell response to fluctuating niche Wnt signaling
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Castillo-Azofeifa, David, Wald, Tomas, Reyes, Efren A, Gallagher, Aaron, Schanin, Julia, Vlachos, Stephanie, Lamarche-Vane, Nathalie, Bomidi, Carolyn, Blutt, Sarah, Estes, Mary K, Nystul, Todd, and Klein, Ophir D
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Biochemistry and Cell Biology ,Biological Sciences ,Cancer ,Colo-Rectal Cancer ,Digestive Diseases ,Regenerative Medicine ,Stem Cell Research ,Stem Cell Research - Nonembryonic - Non-Human ,1.1 Normal biological development and functioning ,Underpinning research ,Aetiology ,2.1 Biological and endogenous factors ,Oral and gastrointestinal ,Animals ,Mice ,Cell Proliferation ,GTPase-Activating Proteins ,Intestinal Mucosa ,Intestines ,Stem Cell Niche ,Stem Cells ,Wnt Signaling Pathway ,Arhgap31 ,Cdc42 ,Dlg1 ,Wnt ,cell death ,epithelium ,intestine ,organoid ,regeneration ,stem cells ,Medical and Health Sciences ,Developmental Biology ,Biological sciences ,Biomedical and clinical sciences - Abstract
A central factor in the maintenance of tissue integrity is the response of stem cells to variations in the levels of niche signals. In the gut, intestinal stem cells (ISCs) depend on Wnt ligands for self-renewal and proliferation. Transient increases in Wnt signaling promote regeneration after injury or in inflammatory bowel diseases, whereas constitutive activation of this pathway leads to colorectal cancer. Here, we report that Discs large 1 (Dlg1), although dispensable for polarity and cellular turnover during intestinal homeostasis, is required for ISC survival in the context of increased Wnt signaling. RNA sequencing (RNA-seq) and genetic mouse models demonstrated that DLG1 regulates the cellular response to increased canonical Wnt ligands. This occurs via the transcriptional regulation of Arhgap31, a GTPase-activating protein that deactivates CDC42, an effector of the non-canonical Wnt pathway. These findings reveal a DLG1-ARHGAP31-CDC42 axis that is essential for the ISC response to increased niche Wnt signaling.
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- 2023
8. Epithelial TNF controls cell differentiation and CFTR activity to maintain intestinal mucin homeostasis
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Reyes, Efren A, Castillo-Azofeifa, David, Rispal, Jérémie, Wald, Tomas, Zwick, Rachel K, Palikuqi, Brisa, Mujukian, Angela, Rabizadeh, Shervin, Gupta, Alexander R, Gardner, James M, Boffelli, Dario, Gartner, Zev J, and Klein, Ophir D
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Biological Sciences ,Medical Physiology ,Biomedical and Clinical Sciences ,Stem Cell Research ,Cystic Fibrosis ,Digestive Diseases ,Autoimmune Disease ,Lung ,Crohn's Disease ,Rare Diseases ,Inflammatory Bowel Disease ,1.1 Normal biological development and functioning ,2.1 Biological and endogenous factors ,Oral and gastrointestinal ,Humans ,Animals ,Mice ,Mucins ,Cystic Fibrosis Transmembrane Conductance Regulator ,Tumor Necrosis Factor Inhibitors ,Epithelial Cells ,Cell Differentiation ,Tumor Necrosis Factors ,Homeostasis ,Cytokines ,Epithelial transport of ions and water ,Gastroenterology ,Molecular genetics ,Medical and Health Sciences ,Immunology ,Biological sciences ,Biomedical and clinical sciences ,Health sciences - Abstract
The gastrointestinal tract relies on the production, maturation, and transit of mucin to protect against pathogens and to lubricate the epithelial lining. Although the molecular and cellular mechanisms that regulate mucin production and movement are beginning to be understood, the upstream epithelial signals that contribute to mucin regulation remain unclear. Here, we report that the inflammatory cytokine tumor necrosis factor (TNF), generated by the epithelium, contributes to mucin homeostasis by regulating both cell differentiation and cystic fibrosis transmembrane conductance regulator (CFTR) activity. We used genetic mouse models and noninflamed samples from patients with inflammatory bowel disease (IBD) undergoing anti-TNF therapy to assess the effect of in vivo perturbation of TNF. We found that inhibition of epithelial TNF promotes the differentiation of secretory progenitor cells into mucus-producing goblet cells. Furthermore, TNF treatment and CFTR inhibition in intestinal organoids demonstrated that TNF promotes ion transport and luminal flow via CFTR. The absence of TNF led to slower gut transit times, which we propose results from increased mucus accumulation coupled with decreased luminal fluid pumping. These findings point to a TNF/CFTR signaling axis in the adult intestine and identify epithelial cell-derived TNF as an upstream regulator of mucin homeostasis.
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- 2023
9. Author Correction: MusMorph, a database of standardized mouse morphology data for morphometric meta-analyses
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Devine, Jay, Vidal-García, Marta, Liu, Wei, Neves, Amanda, Lo Vercio, Lucas D, Green, Rebecca M, Richbourg, Heather A, Marchini, Marta, Unger, Colton M, Nickle, Audrey C, Radford, Bethany, Young, Nathan M, Gonzalez, Paula N, Schuler, Robert E, Bugacov, Alejandro, Rolian, Campbell, Percival, Christopher J, Williams, Trevor, Niswander, Lee, Calof, Anne L, Lander, Arthur D, Visel, Axel, Jirik, Frank R, Cheverud, James M, Klein, Ophir D, Birnbaum, Ramon Y, Merrill, Amy E, Ackermann, Rebecca R, Graf, Daniel, Hemberger, Myriam, Dean, Wendy, Forkert, Nils D, Murray, Stephen A, Westerberg, Henrik, Marcucio, Ralph S, and Hallgrímsson, Benedikt
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Epidemiology ,Biological Sciences ,Health Sciences - Abstract
Correction to: Scientific Data, published online 25 May 2022 The order of the column headings was incorrect in Table 1, meaning values were listed under the wrong descriptions. This has been corrected in the pdf and HTML versions of the article.
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- 2023
10. Intracellular pH dynamics regulates intestinal stem cell lineage specification
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Liu, Yi, Reyes, Efren, Castillo-Azofeifa, David, Klein, Ophir D, Nystul, Todd, and Barber, Diane L
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Biochemistry and Cell Biology ,Biomedical and Clinical Sciences ,Biological Sciences ,Stem Cell Research ,Stem Cell Research - Nonembryonic - Non-Human ,Regenerative Medicine ,Digestive Diseases ,1.1 Normal biological development and functioning ,Mice ,Animals ,Cell Lineage ,Intestines ,Cell Differentiation ,Stem Cells ,Hydrogen-Ion Concentration ,Intestinal Mucosa - Abstract
Intracellular pH dynamics is increasingly recognized to regulate myriad cell behaviors. We report a finding that intracellular pH dynamics also regulates adult stem cell lineage specification. We identify an intracellular pH gradient in mouse small intestinal crypts, lowest in crypt stem cells and increasing along the crypt column. Disrupting this gradient by inhibiting H+ efflux by Na+/H+ exchanger 1 abolishes crypt budding and blocks differentiation of Paneth cells, which are rescued with exogenous WNT. Using single-cell RNA sequencing and lineage tracing we demonstrate that intracellular pH dynamics acts downstream of ATOH1, with increased pH promoting differentiation toward the secretory lineage. Our findings indicate that an increase in pH is required for the lineage specification that contributes to crypt maintenance, establishing a role for intracellular pH dynamics in cell fate decisions within an adult stem cell lineage.
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- 2023
11. Comparing 2D and 3D representations for face-based genetic syndrome diagnosis
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Bannister, Jordan J., Wilms, Matthias, Aponte, J. David, Katz, David C., Klein, Ophir D., Bernier, Francois P., Spritz, Richard A., Hallgrímsson, Benedikt, and Forkert, Nils D.
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- 2023
- Full Text
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12. Cellular shape reinforces niche to stem cell signaling in the small intestine
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Pentinmikko, Nalle, Lozano, Rodrigo, Scharaw, Sandra, Andersson, Simon, Englund, Johanna I, Castillo-Azofeifa, David, Gallagher, Aaron, Broberg, Martin, Song, Ki-Young, Carvajal, Agustín Sola, Speidel, Alessondra T, Sundstrom, Michael, Allbritton, Nancy, Stevens, Molly M, Klein, Ophir D, Teixeira, Ana, and Katajisto, Pekka
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Biochemistry and Cell Biology ,Biomedical and Clinical Sciences ,Biological Sciences ,Stem Cell Research ,Stem Cell Research - Nonembryonic - Non-Human ,Regenerative Medicine ,Digestive Diseases - Abstract
Niche-derived factors regulate tissue stem cells, but apart from the mechanosensory pathways, the effect of niche geometry is not well understood. We used organoids and bioengineered tissue culture platforms to demonstrate that the conical shape of Lgr5+ small intestinal stem cells (ISCs) facilitate their self-renewal and function. Inhibition of non-muscle myosin II (NM II)-driven apical constriction altered ISC shape and reduced niche curvature and stem cell capacity. Niche curvature is decreased in aged mice, suggesting that suboptimal interactions between old ISCs and their niche develop with age. We show that activation of NM IIC or physical restriction to young topology improves in vitro regeneration by old epithelium. We propose that the increase in lateral surface area of ISCs induced by apical constriction promotes interactions between neighboring cells, and the curved topology of the intestinal niche has evolved to maximize signaling between ISCs and neighboring cells.
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- 2022
13. A dominant negative mutation uncovers cooperative control of caudal Wolffian duct development by Sprouty genes
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Altés, Gisela, Vaquero, Marta, Cuesta, Sara, Anerillas, Carlos, Macià, Anna, Espinet, Carme, Ribera, Joan, Bellusci, Saverio, Klein, Ophir D, Yeramian, Andree, Dolcet, Xavi, Egea, Joaquim, and Encinas, Mario
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Biochemistry and Cell Biology ,Biomedical and Clinical Sciences ,Biological Sciences ,Genetics ,Kidney Disease ,Aetiology ,2.1 Biological and endogenous factors ,Animals ,Female ,Male ,Mammals ,Mice ,Mice ,Knockout ,Mutation ,Organogenesis ,Signal Transduction ,Tyrosine ,Wolffian Ducts ,Wolffian duct ,Dominant negative ,Seminal vesicle ,Gartner cyst ,Genitourinary development ,Physiology ,Clinical Sciences ,Biochemistry & Molecular Biology ,Biochemistry and cell biology ,Medical biochemistry and metabolomics ,Oncology and carcinogenesis - Abstract
The Wolffian ducts (WD) are paired epithelial tubules central to the development of the mammalian genitourinary tract. Outgrowths from the WD known as the ureteric buds (UB) generate the collecting ducts of the kidney. Later during development, the caudal portion of the WD will form the vas deferens, epididymis and seminal vesicle in males, and will degenerate in females. While the genetic pathways controlling the development of the UB are firmly established, less is known about those governing development of WD portions caudal to the UB. Sprouty proteins are inhibitors of receptor tyrosine kinase (RTK) signaling in vivo. We have recently shown that homozygous mutation of a conserved tyrosine (Tyr53) of Spry1 results in UB defects indistinguishable from that of Spry1 null mice. Here, we show that heterozygosity for the Spry1 Y53A allele causes caudal WD developmental defects consisting of ectopically branched seminal vesicles in males and persistent WD in females, without affecting kidney development. Detailed analysis reveals that this phenotype also occurs in Spry1+/- mice but with a much lower penetrance, indicating that removal of tyrosine 53 generates a dominant negative mutation in vivo. Supporting this notion, concomitant deletion of one allele of Spry1 and Spry2 also recapitulates the genital phenotype of Spry1Y53A/+ mice with high penetrance. Mechanistically, we show that unlike the effects of Spry1 in kidney development, these caudal WD defects are independent of Ret signaling, but can be completely rescued by lowering the genetic dosage of Fgf10. In conclusion, mutation of tyrosine 53 of Spry1 generates a dominant negative allele that uncovers fine-tuning of caudal WD development by Sprouty genes.
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- 2022
14. Adult dental epithelial stem cell-derived organoids deposit hydroxylapatite biomineral
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Kim, Hyun-Yi, Cooley, Victoria, Kim, Eun-Jung, Li, Shujin, Lee, Jong-Min, Sheyfer, Dina, Liu, Wenjun, Klein, Ophir D., Joester, Derk, and Jung, Han-Sung
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- 2023
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15. A Deep Invertible 3-D Facial Shape Model for Interpretable Genetic Syndrome Diagnosis
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Bannister, Jordan J, Wilms, Matthias, Aponte, J David, Katz, David C, Klein, Ophir D, Bernier, Francois PJ, Spritz, Richard A, Hallgrmsson, Benedikt, and Forkert, Nils D
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Information and Computing Sciences ,Human-Centred Computing ,Bioengineering ,Diagnosis ,Computer-Assisted ,Face ,Humans ,Three-dimensional displays ,Solid modeling ,Surface morphology ,Shape ,Genetics ,Data models ,Morphology ,Genetic syndrome ,normalizing flow ,interpretable machine learning ,3D shape model ,Engineering ,Medical and Health Sciences ,Medical Informatics ,Health services and systems ,Applied computing - Abstract
One of the primary difficulties in treating patients with genetic syndromes is diagnosing their condition. Many syndromes are associated with characteristic facial features that can be imaged and utilized by computer-assisted diagnosis systems. In this work, we develop a novel 3D facial surface modeling approach with the objective of maximizing diagnostic model interpretability within a flexible deep learning framework. Therefore, an invertible normalizing flow architecture is introduced to enable both inferential and generative tasks in a unified and efficient manner. The proposed model can be used (1) to infer syndrome diagnosis and other demographic variables given a 3D facial surface scan and (2) to explain model inferences to non-technical users via multiple interpretability mechanisms. The model was trained and evaluated on more than 4700 facial surface scans from subjects with 47 different syndromes. For the challenging task of predicting syndrome diagnosis given a new 3D facial surface scan, age, and sex of a subject, the model achieves a competitive overall top-1 accuracy of 71%, and a mean sensitivity of 43% across all syndrome classes. We believe that invertible models such as the one presented in this work can achieve competitive inferential performance while greatly increasing model interpretability in the domain of medical diagnosis.
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- 2022
16. Activation of fetal-like molecular programs during regeneration in the intestine and beyond
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Viragova, Sara, Li, Dong, and Klein, Ophir D.
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- 2024
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17. Patterning and folding of intestinal villi by active mesenchymal dewetting
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Huycke, Tyler R., Häkkinen, Teemu J., Miyazaki, Hikaru, Srivastava, Vasudha, Barruet, Emilie, McGinnis, Christopher S., Kalantari, Ali, Cornwall-Scoones, Jake, Vaka, Dedeepya, Zhu, Qin, Jo, Hyunil, Oria, Roger, Weaver, Valerie M., DeGrado, William F., Thomson, Matt, Garikipati, Krishna, Boffelli, Dario, Klein, Ophir D., and Gartner, Zev J.
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- 2024
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18. SRSF1 governs progenitor-specific alternative splicing to maintain adult epithelial tissue homeostasis and renewal
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Yu, Tingsheng, Cazares, Oscar, Tang, Alison D, Kim, Hyun-Yi, Wald, Tomas, Verma, Adya, Liu, Qi, Barcellos-Hoff, Mary Helen, Floor, Stephen N, Jung, Han-Sung, Brooks, Angela N, and Klein, Ophir D
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Biochemistry and Cell Biology ,Biological Sciences ,Genetics ,Stem Cell Research - Nonembryonic - Non-Human ,Stem Cell Research ,Underpinning research ,1.1 Normal biological development and functioning ,Generic health relevance ,Alternative Splicing ,Animals ,Epithelium ,Homeostasis ,Mice ,RNA Splicing ,Serine-Arginine Splicing Factors ,alternative splicing ,incisor ,intestine ,mouse ,progenitor ,splicing factor ,tissue homeostasis ,tissue renewal ,Medical and Health Sciences ,Developmental Biology ,Biochemistry and cell biology - Abstract
Alternative splicing generates distinct mRNA variants and is essential for development, homeostasis, and renewal. Proteins of the serine/arginine (SR)-rich splicing factor family are major splicing regulators that are broadly required for organ development as well as cell and organism viability. However, how these proteins support adult organ function remains largely unknown. Here, we used the continuously growing mouse incisor as a model to dissect the functions of the prototypical SR family protein SRSF1 during tissue homeostasis and renewal. We identified an SRSF1-governed alternative splicing network that is specifically required for dental proliferation and survival of progenitors but dispensable for the viability of differentiated cells. We also observed a similar progenitor-specific role of SRSF1 in the small intestinal epithelium, indicating a conserved function of SRSF1 across adult epithelial tissues. Thus, our findings define a regulatory mechanism by which SRSF1 specifically controls progenitor-specific alternative splicing events to support adult tissue homeostasis and renewal.
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- 2022
19. Microbial signals, MyD88, and lymphotoxin drive TNF-independent intestinal epithelial tissue damage
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Rusu, Iulia, Mennillo, Elvira, Bain, Jared L, Li, Zhongmei, Sun, Xiaofei, Ly, Kimberly M, Rosli, Yenny Y, Naser, Mohammad, Wang, Zunqiu, Advincula, Rommel, Achacoso, Philip, Shao, Ling, Razani, Bahram, Klein, Ophir D, Marson, Alexander, Turnbaugh, Jessie A, Turnbaugh, Peter J, Malynn, Barbara A, Ma, Averil, and Kattah, Michael G
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Digestive Diseases ,Autoimmune Disease ,Crohn's Disease ,Inflammatory Bowel Disease ,Aetiology ,2.1 Biological and endogenous factors ,Oral and gastrointestinal ,Adaptor Proteins ,Signal Transducing ,Animals ,Apoptosis ,Epithelial Cells ,Epithelium ,Humans ,Inflammatory Bowel Diseases ,Intestinal Mucosa ,Lymphotoxin-alpha ,Mice ,Myeloid Differentiation Factor 88 ,Tumor Necrosis Factor Inhibitors ,Apoptosis survival pathways ,Gastroenterology ,Immunology ,Inflammatory bowel disease ,Mouse models ,Medical and Health Sciences - Abstract
Anti-TNF antibodies are effective for treating patients with inflammatory bowel disease (IBD), but many patients fail to respond to anti-TNF therapy, highlighting the importance of TNF-independent disease. We previously demonstrated that acute deletion of 2 IBD susceptibility genes, A20 (Tnfaip3) and Abin-1 (Tnip1), in intestinal epithelial cells (IECs) sensitized mice to both TNF-dependent and TNF-independent death. Here we show that TNF-independent IEC death after A20 and Abin-1 deletion was rescued by germ-free derivation or deletion of MyD88, while deletion of Trif provided only partial protection. Combined deletion of Ripk3 and Casp8, which inhibits both apoptotic and necroptotic death, completely protected against death after acute deletion of A20 and Abin-1 in IECs. A20- and Abin-1-deficient IECs were sensitized to TNF-independent, TNFR1-mediated death in response to lymphotoxin α (LTα) homotrimers. Blockade of LTα in vivo reduced weight loss and improved survival when combined with partial deletion of MyD88. Biopsies of inflamed colon mucosa from patients with IBD exhibited increased LTA and IL1B expression, including a subset of patients with active colitis on anti-TNF therapy. These data show that microbial signals, MyD88, and LTα all contribute to TNF-independent intestinal injury.
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- 2022
20. LGL1 binds to Integrin β1 and inhibits downstream signaling to promote epithelial branching in the mammary gland
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Ma, Rongze, Gong, Difei, You, Huanyang, Xu, Chongshen, Lu, Yunzhe, Bergers, Gabriele, Werb, Zena, Klein, Ophir D, Petritsch, Claudia K, and Lu, Pengfei
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Biochemistry and Cell Biology ,Biological Sciences ,Breast Cancer ,Cancer ,Animals ,Cell Movement ,Cell Polarity ,Cell Proliferation ,Down-Regulation ,Epithelial Cells ,Epithelium ,Female ,Gene Expression Regulation ,Neoplastic ,Glycoproteins ,Integrin beta1 ,Mammary Glands ,Animal ,Mice ,Transgenic ,Models ,Biological ,Morphogenesis ,Protein Binding ,Signal Transduction ,branching morphogenesis ,collective migration ,epithelial migration ,epithelial polarity ,extracellular matrix ,integrin signaling ,tumor-suppressor genes ,Medical Physiology ,Biological sciences - Abstract
Branching morphogenesis is a fundamental process by which organs in invertebrates and vertebrates form branches to expand their surface areas. The current dogma holds that directional cell migration determines where a new branch forms and thus patterns branching. Here, we asked whether mouse Lgl1, a homolog of the Drosophila tumor suppressor Lgl, regulates epithelial polarity in the mammary gland. Surprisingly, mammary glands lacking Lgl1 have normal epithelial polarity, but they form fewer branches. Moreover, we find that Lgl1 null epithelium is unable to directionally migrate, suggesting that migration is not essential for mammary epithelial branching as expected. We show that LGL1 binds to Integrin β1 and inhibits its downstream signaling, and Integrin β1 overexpression blocks epithelial migration, thus recapitulating the Lgl1 null phenotype. Altogether, we demonstrate that Lgl1 modulation of Integrin β1 signaling is essential for directional migration and that epithelial branching in invertebrates and the mammary gland is fundamentally distinct.
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- 2022
21. MusMorph, a database of standardized mouse morphology data for morphometric meta-analyses
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Devine, Jay, Vidal-García, Marta, Liu, Wei, Neves, Amanda, Lo Vercio, Lucas D, Green, Rebecca M, Richbourg, Heather A, Marchini, Marta, Unger, Colton M, Nickle, Audrey C, Radford, Bethany, Young, Nathan M, Gonzalez, Paula N, Schuler, Robert E, Bugacov, Alejandro, Rolian, Campbell, Percival, Christopher J, Williams, Trevor, Niswander, Lee, Calof, Anne L, Lander, Arthur D, Visel, Axel, Jirik, Frank R, Cheverud, James M, Klein, Ophir D, Birnbaum, Ramon Y, Merrill, Amy E, Ackermann, Rebecca R, Graf, Daniel, Hemberger, Myriam, Dean, Wendy, Forkert, Nils D, Murray, Stephen A, Westerberg, Henrik, Marcucio, Ralph S, and Hallgrímsson, Benedikt
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Biological Sciences ,Bioinformatics and Computational Biology ,Bioengineering ,Pediatric ,Genetics ,Animals ,Brain ,Databases ,Factual ,Mice ,X-Ray Microtomography - Abstract
Complex morphological traits are the product of many genes with transient or lasting developmental effects that interact in anatomical context. Mouse models are a key resource for disentangling such effects, because they offer myriad tools for manipulating the genome in a controlled environment. Unfortunately, phenotypic data are often obtained using laboratory-specific protocols, resulting in self-contained datasets that are difficult to relate to one another for larger scale analyses. To enable meta-analyses of morphological variation, particularly in the craniofacial complex and brain, we created MusMorph, a database of standardized mouse morphology data spanning numerous genotypes and developmental stages, including E10.5, E11.5, E14.5, E15.5, E18.5, and adulthood. To standardize data collection, we implemented an atlas-based phenotyping pipeline that combines techniques from image registration, deep learning, and morphometrics. Alongside stage-specific atlases, we provide aligned micro-computed tomography images, dense anatomical landmarks, and segmentations (if available) for each specimen (N = 10,056). Our workflow is open-source to encourage transparency and reproducible data collection. The MusMorph data and scripts are available on FaceBase ( www.facebase.org , https://doi.org/10.25550/3-HXMC ) and GitHub ( https://github.com/jaydevine/MusMorph ).
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- 2022
22. CNPY4 inhibits the Hedgehog pathway by modulating membrane sterol lipids
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Lo, Megan, Sharir, Amnon, Paul, Michael D, Torosyan, Hayarpi, Agnew, Christopher, Li, Amy, Neben, Cynthia, Marangoni, Pauline, Xu, Libin, Raleigh, David R, Jura, Natalia, and Klein, Ophir D
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Biochemistry and Cell Biology ,Biological Sciences ,1.1 Normal biological development and functioning ,Underpinning research ,Generic health relevance ,Cholesterol ,Hedgehog Proteins ,Membrane Lipids ,Signal Transduction ,Sterols - Abstract
The Hedgehog (HH) pathway is critical for development and adult tissue homeostasis. Aberrant HH signaling can lead to congenital malformations and diseases including cancer. Although cholesterol and several oxysterol lipids have been shown to play crucial roles in HH activation, the molecular mechanisms governing their regulation remain unresolved. Here, we identify Canopy4 (CNPY4), a Saposin-like protein, as a regulator of the HH pathway that modulates levels of membrane sterol lipids. Cnpy4-/- embryos exhibit multiple defects consistent with HH signaling perturbations, most notably changes in digit number. Knockdown of Cnpy4 hyperactivates the HH pathway in vitro and elevates membrane levels of accessible sterol lipids, such as cholesterol, an endogenous ligand involved in HH activation. Our data demonstrate that CNPY4 is a negative regulator that fine-tunes HH signal transduction, revealing a previously undescribed facet of HH pathway regulation that operates through control of membrane composition.
- Published
- 2022
23. An interactive atlas of three-dimensional syndromic facial morphology
- Author
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Aponte, J. David, Bannister, Jordan J., Hoskens, Hanne, Matthews, Harold, Katsura, Kaitlin, Da Silva, Cassidy, Cruz, Tim, Pilz, Julie H.M., Spritz, Richard A., Forkert, Nils D., Claes, Peter, Bernier, Francois P., Klein, Ophir D., Katz, David C., and Hallgrímsson, Benedikt
- Published
- 2024
- Full Text
- View/download PDF
24. Application of full-genome analysis to diagnose rare monogenic disorders.
- Author
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Shieh, Joseph T, Penon-Portmann, Monica, Wong, Karen HY, Levy-Sakin, Michal, Verghese, Michelle, Slavotinek, Anne, Gallagher, Renata C, Mendelsohn, Bryce A, Tenney, Jessica, Beleford, Daniah, Perry, Hazel, Chow, Stephen K, Sharo, Andrew G, Brenner, Steven E, Qi, Zhongxia, Yu, Jingwei, Klein, Ophir D, Martin, David, Kwok, Pui-Yan, and Boffelli, Dario
- Subjects
Human Genome ,Genetics ,Genetic Testing ,Prevention ,4.1 Discovery and preclinical testing of markers and technologies ,4.2 Evaluation of markers and technologies - Abstract
Current genetic testenhancer and narrows the diagnostic intervals for rare diseases provide a diagnosis in only a modest proportion of cases. The Full-Genome Analysis method, FGA, combines long-range assembly and whole-genome sequencing to detect small variants, structural variants with breakpoint resolution, and phasing. We built a variant prioritization pipeline and tested FGA's utility for diagnosis of rare diseases in a clinical setting. FGA identified structural variants and small variants with an overall diagnostic yield of 40% (20 of 50 cases) and 35% in exome-negative cases (8 of 23 cases), 4 of these were structural variants. FGA detected and mapped structural variants that are missed by short reads, including non-coding duplication, and phased variants across long distances of more than 180 kb. With the prioritization algorithm, longer DNA technologies could replace multiple tests for monogenic disorders and expand the range of variants detected. Our study suggests that genomes produced from technologies like FGA can improve variant detection and provide higher resolution genome maps for future application.
- Published
- 2021
25. Inflation-collapse dynamics drive patterning and morphogenesis in intestinal organoids
- Author
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Tallapragada, Naren P, Cambra, Hailey M, Wald, Tomas, Keough Jalbert, Samantha, Abraham, Diana M, Klein, Ophir D, and Klein, Allon M
- Subjects
Biological Sciences ,Biomedical and Clinical Sciences ,Regenerative Medicine ,Stem Cell Research ,Digestive Diseases ,1.1 Normal biological development and functioning ,Underpinning research ,Cell Differentiation ,Intestinal Mucosa ,Intestines ,Morphogenesis ,Organoids ,Stem Cells ,intestinal stem cells ,live imaging ,mechanobiology ,organoids ,self-organization ,size control ,stretch-responsive cell ,Medical and Health Sciences ,Developmental Biology ,Biological sciences ,Biomedical and clinical sciences - Abstract
How stem cells self-organize to form structured tissues is an unsolved problem. Intestinal organoids offer a model of self-organization as they generate stem cell zones (SCZs) of typical size even without a spatially structured environment. Here we examine processes governing the size of SCZs. We improve the viability and homogeneity of intestinal organoid cultures to enable long-term time-lapse imaging of multiple organoids in parallel. We find that SCZs are shaped by fission events under strong control of ion channel-mediated inflation and mechanosensitive Piezo-family channels. Fission occurs through stereotyped modes of dynamic behavior that differ in their coordination of budding and differentiation. Imaging and single-cell transcriptomics show that inflation drives acute stem cell differentiation and induces a stretch-responsive cell state characterized by large transcriptional changes, including upregulation of Piezo1. Our results reveal an intrinsic capacity of the intestinal epithelium to self-organize by modulating and then responding to its mechanical state.
- Published
- 2021
26. Bcl-6 expression by CD4+ T cells determines concomitant immunity and host resistance across distinct parasitic infections
- Author
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Meli, Alexandre P., Russell, Gabriel A., Swaminathan, Sharada, Weichselbaum, Laura, MacMahon, Clara A., Pernet, Erwan, Karo-Atar, Danielle, Rogers, Dakota, Rochette, Annie, Fontes, Ghislaine, Mandl, Judith N., Divangahi, Maziar, Klein, Ophir D., Gregorieff, Alex, Stäger, Simona, and King, Irah L.
- Published
- 2023
- Full Text
- View/download PDF
27. Large-scale open-source three-dimensional growth curves for clinical facial assessment and objective description of facial dysmorphism.
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Matthews, Harold S, Palmer, Richard L, Baynam, Gareth S, Quarrell, Oliver W, Klein, Ophir D, Spritz, Richard A, Hennekam, Raoul C, Walsh, Susan, Shriver, Mark, Weinberg, Seth M, Hallgrimsson, Benedikt, Hammond, Peter, Penington, Anthony J, Peeters, Hilde, and Claes, Peter D
- Abstract
Craniofacial dysmorphism is associated with thousands of genetic and environmental disorders. Delineation of salient facial characteristics can guide clinicians towards a correct clinical diagnosis and understanding the pathogenesis of the disorder. Abnormal facial shape might require craniofacial surgical intervention, with the restoration of normal shape an important surgical outcome. Facial anthropometric growth curves or standards of single inter-landmark measurements have traditionally supported assessments of normal and abnormal facial shape, for both clinical and research applications. However, these fail to capture the full complexity of facial shape. With the increasing availability of 3D photographs, methods of assessment that take advantage of the rich information contained in such images are needed. In this article we derive and present open-source three-dimensional (3D) growth curves of the human face. These are sequences of age and sex-specific expected 3D facial shapes and statistical models of the variation around the expected shape, derived from 5443 3D images. We demonstrate the use of these growth curves for assessing patients and show that they identify normal and abnormal facial morphology independent from age-specific facial features. 3D growth curves can facilitate use of state-of-the-art 3D facial shape assessment by the broader clinical and biomedical research community. This advance in phenotype description will support clinical diagnosis and the understanding of disease pathogenesis including genotype-phenotype relations.
- Published
- 2021
28. Multiple endocrine defects in adult-onset Sprouty1/2/4 triple knockout mice
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Altés, Gisela, Olomí, Anna, Perramon-Güell, Aida, Hernández, Sara, Casanovas, Anna, Pérez, Aurora, Díaz-Tocados, Juan Miguel, Valdivielso, José Manuel, Megino, Cristina, Navaridas, Raúl, Matias-Guiu, Xavier, Klein, Ophir D., Egea, Joaquim, Dolcet, Xavi, Yeramian, Andrée, and Encinas, Mario
- Published
- 2024
- Full Text
- View/download PDF
29. Myosin II mediates Shh signals to shape dental epithelia via control of cell adhesion and movement
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Du, Wei, primary, Verma, Adya, additional, Ye, Qianlin, additional, Du, Wen, additional, Lin, Sandy, additional, Yamanaka, Atsushi, additional, Klein, Ophir D., additional, and Hu, Jimmy K., additional
- Published
- 2024
- Full Text
- View/download PDF
30. Mapping genes for human face shape: exploration of univariate phenotyping strategies
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Yuan, Meng, primary, Goovaerts, Seppe, additional, Vanneste, Michiel, additional, Matthews, Harold, additional, Hoskens, Hanne, additional, Richmond, Stephen, additional, Klein, Ophir D, additional, Spritz, Richard A, additional, Hallgrimsson, Benedikt, additional, Walsh, Susan, additional, Shriver, Mark D, additional, Shaffer, John R, additional, Weinberg, Seth M, additional, Peeters, Hilde, additional, and Claes, Peter, additional
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- 2024
- Full Text
- View/download PDF
31. Transit-Amplifying Cells Coordinate Changes in Intestinal Epithelial Cell-Type Composition
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Sanman, Laura E, Chen, Ina W, Bieber, Jake M, Steri, Veronica, Trentesaux, Coralie, Hann, Byron, Klein, Ophir D, Wu, Lani F, and Altschuler, Steven J
- Subjects
Biochemistry and Cell Biology ,Biological Sciences ,Stem Cell Research - Nonembryonic - Non-Human ,Stem Cell Research ,Digestive Diseases ,1.1 Normal biological development and functioning ,Underpinning research ,Aetiology ,2.1 Biological and endogenous factors ,Animals ,Cell Proliferation ,Epithelial Cells ,ErbB Receptors ,Humans ,Interleukin-4 ,Intestinal Absorption ,Intestines ,Male ,Mice ,Inbred C57BL ,Models ,Biological ,Organoids ,Protein Interaction Mapping ,Signal Transduction ,cell-type composition ,high-throughput platform ,intestinal epithelium ,lineage model ,organoid ,systems biology ,Medical and Health Sciences ,Developmental Biology ,Biochemistry and cell biology - Abstract
Renewing tissues have the remarkable ability to continually produce both proliferative progenitor and specialized differentiated cell types. How are complex milieus of microenvironmental signals interpreted to coordinate tissue-cell-type composition? Here, we investigate the responses of intestinal epithelium to individual and paired perturbations across eight epithelial signaling pathways. Using a high-throughput approach that combines enteroid monolayers and quantitative imaging, we identified conditions that enrich for specific cell types as well as interactions between pathways. Importantly, we found that modulation of transit-amplifying cell proliferation changes the ratio of differentiated secretory to absorptive cell types. These observations highlight an underappreciated role for transit-amplifying cells in the tuning of differentiated cell-type composition.
- Published
- 2021
32. LGR5 in breast cancer and ductal carcinoma in situ: a diagnostic and prognostic biomarker and a therapeutic target
- Author
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Hagerling, Catharina, Owyong, Mark, Sitarama, Vaishnavi, Wang, Chih-Yang, Lin, Charlene, van den Bijgaart, Renske JE, Koopman, Charlotte D, Brenot, Audrey, Nanjaraj, Ankitha, Wärnberg, Fredrik, Jirström, Karin, Klein, Ophir D, Werb, Zena, and Plaks, Vicki
- Subjects
Biomedical and Clinical Sciences ,Oncology and Carcinogenesis ,Immunology ,Breast Cancer ,Biotechnology ,Prevention ,Cancer ,Adult ,Aged ,Aged ,80 and over ,Animals ,Biomarkers ,Tumor ,Breast Neoplasms ,Carcinoma ,Intraductal ,Noninfiltrating ,Cell Line ,Tumor ,Female ,Heterografts ,Humans ,Mice ,Middle Aged ,Prognosis ,RNA ,Neoplasm ,Real-Time Polymerase Chain Reaction ,Receptor ,ErbB-2 ,Receptors ,G-Protein-Coupled ,Tissue Array Analysis ,LGR5 ,Breast cancer ,DCIS ,Estrogen receptor ,Targeted therapy ,Receptor ,erbB-2 ,Public Health and Health Services ,Oncology & Carcinogenesis ,Oncology and carcinogenesis ,Epidemiology - Abstract
BackgroundNovel biomarkers are required to discern between breast tumors that should be targeted for treatment from those that would never become clinically apparent and/or life threatening for patients. Moreover, therapeutics that specifically target breast cancer (BC) cells with tumor-initiating capacity to prevent recurrence are an unmet need. We investigated the clinical importance of LGR5 in BC and ductal carcinoma in situ (DCIS) to explore LGR5 as a biomarker and a therapeutic target.MethodsWe stained BC (n = 401) and DCIS (n = 119) tissue microarrays with an antibody against LGR5. We examined an LGR5 knockdown ER- cell line that was orthotopically transplanted and used for in vitro colony assays. We also determined the tumor-initiating role of Lgr5 in lineage-tracing experiments. Lastly, we transplanted ER- patient-derived xenografts into mice that were subsequently treated with a LGR5 antibody drug conjugate (anti-LGR5-ADC).ResultsLGR5 expression correlated with small tumor size, lower grade, lymph node negativity, and ER-positivity. ER+ patients with LGR5high tumors rarely had recurrence, while high-grade ER- patients with LGR5high expression recurred and died due to BC more often. Intriguingly, all the DCIS patients who later died of BC had LGR5-positive tumors. Colony assays and xenograft experiments substantiated a role for LGR5 in ER- tumor initiation and subsequent growth, which was further validated by lineage-tracing experiments in ER- /triple-negative BC mouse models. Importantly, by utilizing LGR5high patient-derived xenografts, we showed that anti-LGR5-ADC should be considered as a therapeutic for high-grade ER- BC.ConclusionLGR5 has distinct roles in ER- vs. ER+ BC with potential clinical applicability as a biomarker to identify patients in need of therapy and could serve as a therapeutic target for high-grade ER- BC.
- Published
- 2020
33. KrasP34R and KrasT58I mutations induce distinct RASopathy phenotypes in mice.
- Author
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Wong, Jasmine C, Perez-Mancera, Pedro A, Huang, Tannie Q, Kim, Jangkyung, Grego-Bessa, Joaquim, Del Pilar Alzamora, Maria, Kogan, Scott C, Sharir, Amnon, Keefe, Susan H, Morales, Carolina E, Schanze, Denny, Castel, Pau, Hirose, Kentaro, Huang, Guo N, Zenker, Martin, Sheppard, Dean, Klein, Ophir D, Tuveson, David A, Braun, Benjamin S, and Shannon, Kevin
- Subjects
Animals ,Mice ,Inbred C57BL ,Mice ,Craniosynostoses ,Lung Diseases ,Cardiomyopathies ,Hematologic Diseases ,Pregnancy ,Mutation ,Female ,Male ,Proto-Oncogene Proteins p21(ras) ,Genetic diseases ,Genetics ,Mouse models ,Signal transduction - Abstract
Somatic KRAS mutations are highly prevalent in many cancers. In addition, a distinct spectrum of germline KRAS mutations causes developmental disorders called RASopathies. The mutant proteins encoded by these germline KRAS mutations are less biochemically and functionally activated than those in cancer. We generated mice harboring conditional KrasLSL-P34Rand KrasLSL-T58I knock-in alleles and characterized the consequences of each mutation in vivo. Embryonic expression of KrasT58I resulted in craniofacial abnormalities reminiscent of those seen in RASopathy disorders, and these mice exhibited hyperplastic growth of multiple organs, modest alterations in cardiac valvulogenesis, myocardial hypertrophy, and myeloproliferation. By contrast, embryonic KrasP34R expression resulted in early perinatal lethality from respiratory failure due to defective lung sacculation, which was associated with aberrant ERK activity in lung epithelial cells. Somatic Mx1-Cre-mediated activation in the hematopoietic compartment showed that KrasP34R and KrasT58I expression had distinct signaling effects, despite causing a similar spectrum of hematologic diseases. These potentially novel strains are robust models for investigating the consequences of expressing endogenous levels of hyperactive K-Ras in different developing and adult tissues, for comparing how oncogenic and germline K-Ras proteins perturb signaling networks and cell fate decisions, and for performing preclinical therapeutic trials.
- Published
- 2020
34. Engineering synthetic morphogen systems that can program multicellular patterning
- Author
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Toda, Satoshi, McKeithan, Wesley L, Hakkinen, Teemu J, Lopez, Pilar, Klein, Ophir D, and Lim, Wendell A
- Subjects
Biochemistry and Cell Biology ,Engineering ,Biological Sciences ,Underpinning research ,1.1 Normal biological development and functioning ,Animals ,Body Patterning ,Drosophila melanogaster ,Fibroblasts ,Green Fluorescent Proteins ,Protein Engineering ,Receptors ,Notch ,Tissue Engineering ,General Science & Technology - Abstract
In metazoan tissues, cells decide their fates by sensing positional information provided by specialized morphogen proteins. To explore what features are sufficient for positional encoding, we asked whether arbitrary molecules (e.g., green fluorescent protein or mCherry) could be converted into synthetic morphogens. Synthetic morphogens expressed from a localized source formed a gradient when trapped by surface-anchoring proteins, and they could be sensed by synthetic receptors. Despite their simplicity, these morphogen systems yielded patterns reminiscent of those observed in vivo. Gradients could be reshaped by altering anchor density or by providing a source of competing inhibitor. Gradient interpretation could be altered by adding feedback loops or morphogen cascades to receiver cell response circuits. Orthogonal cell-cell communication systems provide insight into morphogen evolution and a platform for engineering tissues.
- Published
- 2020
35. Safety and immunogenicity of Fc‐EDA, a recombinant ectodysplasin A1 replacement protein, in human subjects
- Author
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Körber, Iris, Klein, Ophir D, Morhart, Patrick, Faschingbauer, Florian, Grange, Dorothy K, Clarke, Angus, Bodemer, Christine, Maitz, Silvia, Huttner, Kenneth, Kirby, Neil, Durand, Caroline, and Schneider, Holm
- Subjects
Biomedical and Clinical Sciences ,Dentistry ,Clinical Research ,Pediatric Research Initiative ,Perinatal Period - Conditions Originating in Perinatal Period ,Infant Mortality ,Pediatric ,6.1 Pharmaceuticals ,Evaluation of treatments and therapeutic interventions ,Good Health and Well Being ,Adult ,Animals ,Ectodysplasins ,Female ,Humans ,Immunoglobulin Fc Fragments ,Infant ,Infant ,Newborn ,Male ,Pregnancy ,Recombinant Fusion Proteins ,Research Subjects ,drug safety ,ectodermal dysplasia ,ectodysplasin A ,immunogenicity ,prenatal therapy ,protein replacement ,Pharmacology and Pharmaceutical Sciences ,Pharmacology & Pharmacy ,Pharmacology and pharmaceutical sciences - Abstract
In X-linked hypohidrotic ectodermal dysplasia, the most frequent ectodermal dysplasia, an inherited deficiency of the signalling protein ectodysplasin A1 (EDA1) impairs the development of the skin and its appendages, various eccrine glands, and dentition. The severe hypohidrosis common to X-linked hypohidrotic ectodermal dysplasia patients may lead to life-threatening hyperthermia, especially during hot weather or febrile illness. Fc-EDA, an EDA1 replacement protein known to prevent the disease in newborn animals, was tested in 2 clinical trials (human adults and neonates) and additionally administered under compassionate use to 3 infants in utero. The data support the safety of Fc-EDA and efficacy if applied prenatally. Anti-drug antibodies were detected after intravenous administration in adult males and nonpregnant females, but not in pregnant women when Fc-EDA was delivered intra-amniotically. Most importantly, there was no detectable immune response to the investigational drug in neonates treated by intravenous infusions and in infants who had received Fc-EDA in utero. In conclusion, the safety profile of this drug encourages further development of prenatal EDA1 replacement therapy.
- Published
- 2020
36. Automated syndrome diagnosis by three-dimensional facial imaging
- Author
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Hallgrímsson, Benedikt, Aponte, J David, Katz, David C, Bannister, Jordan J, Riccardi, Sheri L, Mahasuwan, Nick, McInnes, Brenda L, Ferrara, Tracey M, Lipman, Danika M, Neves, Amanda B, Spitzmacher, Jared AJ, Larson, Jacinda R, Bellus, Gary A, Pham, Anh M, Aboujaoude, Elias, Benke, Timothy A, Chatfield, Kathryn C, Davis, Shanlee M, Elias, Ellen R, Enzenauer, Robert W, French, Brooke M, Pickler, Laura L, Shieh, Joseph TC, Slavotinek, Anne, Harrop, A Robertson, Innes, A Micheil, McCandless, Shawn E, McCourt, Emily A, Meeks, Naomi JL, Tartaglia, Nicole R, Tsai, Anne C-H, Wyse, J Patrick H, Bernstein, Jonathan A, Sanchez-Lara, Pedro A, Forkert, Nils D, Bernier, Francois P, Spritz, Richard A, and Klein, Ophir D
- Subjects
Biological Sciences ,Genetics ,Clinical Research ,2.1 Biological and endogenous factors ,Aetiology ,Congenital ,Face ,Humans ,Imaging ,Three-Dimensional ,Syndrome ,syndromes ,facial imaging ,deep phenotyping ,diagnosis ,morphometrics ,Clinical Sciences ,Genetics & Heredity - Abstract
PurposeDeep phenotyping is an emerging trend in precision medicine for genetic disease. The shape of the face is affected in 30-40% of known genetic syndromes. Here, we determine whether syndromes can be diagnosed from 3D images of human faces.MethodsWe analyzed variation in three-dimensional (3D) facial images of 7057 subjects: 3327 with 396 different syndromes, 727 of their relatives, and 3003 unrelated, unaffected subjects. We developed and tested machine learning and parametric approaches to automated syndrome diagnosis using 3D facial images.ResultsUnrelated, unaffected subjects were correctly classified with 96% accuracy. Considering both syndromic and unrelated, unaffected subjects together, balanced accuracy was 73% and mean sensitivity 49%. Excluding unrelated, unaffected subjects substantially improved both balanced accuracy (78.1%) and sensitivity (56.9%) of syndrome diagnosis. The best predictors of classification accuracy were phenotypic severity and facial distinctiveness of syndromes. Surprisingly, unaffected relatives of syndromic subjects were frequently classified as syndromic, often to the syndrome of their affected relative.ConclusionDeep phenotyping by quantitative 3D facial imaging has considerable potential to facilitate syndrome diagnosis. Furthermore, 3D facial imaging of "unaffected" relatives may identify unrecognized cases or may reveal novel examples of semidominant inheritance.
- Published
- 2020
37. Asymmetric Stratification-Induced Polarity Loss and Coordinated Individual Cell Movements Drive Directional Migration of Vertebrate Epithelium
- Author
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Lu, Yunzhe, Deng, Ruolan, You, Huanyang, Xu, Yishu, Antos, Christopher, Sun, Jianlong, Klein, Ophir D, and Lu, Pengfei
- Subjects
Biochemistry and Cell Biology ,Biological Sciences ,1.1 Normal biological development and functioning ,Animals ,Cell Movement ,Cell Polarity ,Cell Proliferation ,Cell Surface Extensions ,Dogs ,Epithelial Cells ,Epithelium ,Female ,Fibroblast Growth Factor 10 ,Green Fluorescent Proteins ,Madin Darby Canine Kidney Cells ,Mammary Glands ,Animal ,Mice ,Organoids ,Signal Transduction ,Vertebrates ,FGF gradient ,apicobasal polarity ,branching morphogenesis ,cell polarity ,chemotaxis ,collective migration ,epithelial polarity ,epithelial stratification ,epithelial-mesenchymal transition ,front-rear polarity ,Medical Physiology ,Biological sciences - Abstract
Collective migration is essential for development, wound repair, and cancer metastasis. For most collective systems, "leader cells" determine both the direction and the power of the migration. It has remained unclear, however, how the highly polarized vertebrate epithelium migrates directionally during branching morphogenesis. We show here that, unlike in other systems, front-rear polarity of the mammary epithelium is set up by preferential cell proliferation in the front in response to the FGF10 gradient. This leads to frontal stratification, loss of apicobasal polarity, and leader cell formation. Leader cells are a dynamic population and move faster and more directionally toward the FGF10 signal than do follower cells, partly because of their intraepithelial protrusions toward the signal. Together, our data show that directional migration of the mammary epithelium is a unique multistep process and that, despite sharing remarkable cellular and molecular similarities, vertebrate and invertebrate epithelial branching are fundamentally distinct processes.
- Published
- 2020
38. Dental cell type atlas reveals stem and differentiated cell types in mouse and human teeth.
- Author
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Krivanek, Jan, Soldatov, Ruslan A, Kastriti, Maria Eleni, Chontorotzea, Tatiana, Herdina, Anna Nele, Petersen, Julian, Szarowska, Bara, Landova, Marie, Matejova, Veronika Kovar, Holla, Lydie Izakovicova, Kuchler, Ulrike, Zdrilic, Ivana Vidovic, Vijaykumar, Anushree, Balic, Anamaria, Marangoni, Pauline, Klein, Ophir D, Neves, Vitor CM, Yianni, Val, Sharpe, Paul T, Harkany, Tibor, Metscher, Brian D, Bajénoff, Marc, Mina, Mina, Fried, Kaj, Kharchenko, Peter V, and Adameyko, Igor
- Subjects
Odontoblasts ,Epithelial Cells ,Stem Cells ,Tooth ,Incisor ,Molar ,Mesoderm ,Animals ,Mice ,Inbred C57BL ,Humans ,Mice ,Models ,Animal ,Cell Differentiation ,Gene Expression Regulation ,Developmental ,Genetic Heterogeneity ,Adolescent ,Adult ,Female ,Male ,Young Adult - Abstract
Understanding cell types and mechanisms of dental growth is essential for reconstruction and engineering of teeth. Therefore, we investigated cellular composition of growing and non-growing mouse and human teeth. As a result, we report an unappreciated cellular complexity of the continuously-growing mouse incisor, which suggests a coherent model of cell dynamics enabling unarrested growth. This model relies on spatially-restricted stem, progenitor and differentiated populations in the epithelial and mesenchymal compartments underlying the coordinated expansion of two major branches of pulpal cells and diverse epithelial subtypes. Further comparisons of human and mouse teeth yield both parallelisms and differences in tissue heterogeneity and highlight the specifics behind growing and non-growing modes. Despite being similar at a coarse level, mouse and human teeth reveal molecular differences and species-specific cell subtypes suggesting possible evolutionary divergence. Overall, here we provide an atlas of human and mouse teeth with a focus on growth and differentiation.
- Published
- 2020
39. FaceBase 3: analytical tools and FAIR resources for craniofacial and dental research.
- Author
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Samuels, Bridget D, Aho, Robert, Brinkley, James F, Bugacov, Alejandro, Feingold, Eleanor, Fisher, Shannon, Gonzalez-Reiche, Ana S, Hacia, Joseph G, Hallgrimsson, Benedikt, Hansen, Karissa, Harris, Matthew P, Ho, Thach-Vu, Holmes, Greg, Hooper, Joan E, Jabs, Ethylin Wang, Jones, Kenneth L, Kesselman, Carl, Klein, Ophir D, Leslie, Elizabeth J, Li, Hong, Liao, Eric C, Long, Hannah, Lu, Na, Maas, Richard L, Marazita, Mary L, Mohammed, Jaaved, Prescott, Sara, Schuler, Robert, Selleri, Licia, Spritz, Richard A, Swigut, Tomek, van Bakel, Harm, Visel, Axel, Welsh, Ian, Williams, Cristina, Williams, Trevor J, Wysocka, Joanna, Yuan, Yuan, and Chai, Yang
- Subjects
Skull ,Facial Bones ,Animals ,Humans ,Reproducibility of Results ,Dental Research ,Databases ,Factual ,Research Personnel ,Craniofacial development ,Data resource ,FaceBase Consortium ,Human ,Mouse ,Zebrafish ,Databases ,Factual ,Biological Sciences ,Medical and Health Sciences - Abstract
The FaceBase Consortium was established by the National Institute of Dental and Craniofacial Research in 2009 as a 'big data' resource for the craniofacial research community. Over the past decade, researchers have deposited hundreds of annotated and curated datasets on both normal and disordered craniofacial development in FaceBase, all freely available to the research community on the FaceBase Hub website. The Hub has developed numerous visualization and analysis tools designed to promote integration of multidisciplinary data while remaining dedicated to the FAIR principles of data management (findability, accessibility, interoperability and reusability) and providing a faceted search infrastructure for locating desired data efficiently. Summaries of the datasets generated by the FaceBase projects from 2014 to 2019 are provided here. FaceBase 3 now welcomes contributions of data on craniofacial and dental development in humans, model organisms and cell lines. Collectively, the FaceBase Consortium, along with other NIH-supported data resources, provide a continuously growing, dynamic and current resource for the scientific community while improving data reproducibility and fulfilling data sharing requirements.
- Published
- 2020
40. Fully Automatic Landmarking of Syndromic 3D Facial Surface Scans Using 2D Images.
- Author
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Bannister, Jordan J, Crites, Sebastian R, Aponte, J David, Katz, David C, Wilms, Matthias, Klein, Ophir D, Bernier, Francois PJ, Spritz, Richard A, Hallgrímsson, Benedikt, and Forkert, Nils D
- Subjects
3D surface scan ,facial landmarking ,genetic syndrome ,Environmental Science and Management ,Ecology ,Analytical Chemistry ,Distributed Computing ,Electrical and Electronic Engineering - Abstract
3D facial landmarks are known to be diagnostically relevant biometrics for many genetic syndromes. The objective of this study was to extend a state-of-the-art image-based 2D facial landmarking algorithm for the challenging task of 3D landmark identification on subjects with genetic syndromes, who often have moderate to severe facial dysmorphia. The automatic 3D facial landmarking algorithm presented here uses 2D image-based facial detection and landmarking models to identify 12 landmarks on 3D facial surface scans. The landmarking algorithm was evaluated using a test set of 444 facial scans with ground truth landmarks identified by two different human observers. Three hundred and sixty nine of the subjects in the test set had a genetic syndrome that is associated with facial dysmorphology. For comparison purposes, the manual landmarks were also used to initialize a non-linear surface-based registration of a non-syndromic atlas to each subject scan. Compared to the average intra- and inter-observer landmark distances of 1.1 mm and 1.5 mm respectively, the average distance between the manual landmark positions and those produced by the automatic image-based landmarking algorithm was 2.5 mm. The average error of the registration-based approach was 3.1 mm. Comparing the distributions of Procrustes distances from the mean for each landmarking approach showed that the surface registration algorithm produces a systemic bias towards the atlas shape. In summary, the image-based automatic landmarking approach performed well on this challenging test set, outperforming a semi-automatic surface registration approach, and producing landmark errors that are comparable to state-of-the-art 3D geometry-based facial landmarking algorithms evaluated on non-syndromic subjects.
- Published
- 2020
41. Tools and Concepts for Interrogating and Defining Cellular Identity
- Author
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McKinley, Kara L, Castillo-Azofeifa, David, and Klein, Ophir D
- Subjects
Biochemistry and Cell Biology ,Biological Sciences ,Underpinning research ,1.1 Normal biological development and functioning ,Generic health relevance ,Homeostasis ,Single-Cell Analysis ,Stem Cells ,cell competition ,cell type ,microscopy ,organoid ,plasticity ,scRNA-seq ,smFISH ,stem cell ,Medical and Health Sciences ,Developmental Biology ,Biological sciences ,Biomedical and clinical sciences - Abstract
Defining the mechanisms that generate specialized cell types and coordinate their functions is critical for understanding organ development and renewal. New tools and discoveries are challenging and refining our definitions of a cell type. A rapidly growing toolkit for single-cell analyses has expanded the number of markers that can be assigned to a cell simultaneously, revealing heterogeneity within cell types that were previously regarded as homogeneous populations. Additionally, cell types defined by specific molecular markers can exhibit distinct, context-dependent functions; for example, between tissues in homeostasis and those responding to damage. Here we review the current technologies used to identify and characterize cells, and we discuss how experimental and pathological perturbations are adding increasing complexity to our definitions of cell identity.
- Published
- 2020
42. Lgr5+ telocytes are a signaling source at the intestinal villus tip.
- Author
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Bahar Halpern, Keren, Massalha, Hassan, Zwick, Rachel K, Moor, Andreas E, Castillo-Azofeifa, David, Rozenberg, Milena, Farack, Lydia, Egozi, Adi, Miller, Dan R, Averbukh, Inna, Harnik, Yotam, Weinberg-Corem, Noa, de Sauvage, Frederic J, Amit, Ido, Klein, Ophir D, Shoshkes-Carmel, Michal, and Itzkovitz, Shalev
- Subjects
Intestinal Mucosa ,Enterocytes ,Intestine ,Small ,Stromal Cells ,Animals ,Mice ,Inbred C57BL ,Mice ,Receptors ,G-Protein-Coupled ,Male ,Wnt-5a Protein ,Intestine ,Small ,Inbred C57BL ,Receptors ,G-Protein-Coupled - Abstract
The intestinal epithelium is a structured organ composed of crypts harboring Lgr5+ stem cells, and villi harboring differentiated cells. Spatial transcriptomics have demonstrated profound zonation of epithelial gene expression along the villus axis, but the mechanisms shaping this spatial variability are unknown. Here, we combine laser capture micro-dissection and single cell RNA sequencing to uncover spatially zonated populations of mesenchymal cells along the crypt-villus axis. These include villus tip telocytes (VTTs) that express Lgr5, a gene previously considered a specific crypt epithelial stem cell marker. VTTs are elongated cells that line the villus tip epithelium and signal through Bmp morphogens and the non-canonical Wnt5a ligand. Their ablation is associated with perturbed zonation of enterocyte genes induced at the villus tip. Our study provides a spatially-resolved cell atlas of the small intestinal stroma and exposes Lgr5+ villus tip telocytes as regulators of the epithelial spatial expression programs along the villus axis.
- Published
- 2020
43. From gut to glutes: The critical role of niche signals in the maintenance and renewal of adult stem cells
- Author
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Trentesaux, Coralie, Striedinger, Katharine, Pomerantz, Jason H, and Klein, Ophir D
- Subjects
Biochemistry and Cell Biology ,Biological Sciences ,Stem Cell Research ,Stem Cell Research - Nonembryonic - Non-Human ,Regenerative Medicine ,1.1 Normal biological development and functioning ,Underpinning research ,Adult Stem Cells ,Cell Differentiation ,Gastrointestinal Microbiome ,Humans ,Signal Transduction ,Stem Cell Niche ,Stem cells ,Stem cell niche ,Intestinal stem cells ,Muscle stem cells ,Satellite cells ,Cell signaling ,Muscle stem cells Satellite cells ,Developmental Biology ,Biochemistry and cell biology - Abstract
Stem cell behavior is tightly regulated by spatiotemporal signaling from the niche, which is a four-dimensional microenvironment that can instruct stem cells to remain quiescent, self-renew, proliferate, or differentiate. In this review, we discuss recent advances in understanding the signaling cues provided by the stem cell niche in two contrasting adult tissues, the rapidly cycling intestinal epithelium and the slowly renewing skeletal muscle. Drawing comparisons between these two systems, we discuss the effects of niche-derived growth factors and signaling molecules, metabolic cues, the extracellular matrix and biomechanical cues, and immune signals on stem cells. We also discuss the influence of the niche in defining stem cell identity and function in both normal and pathophysiologic states.
- Published
- 2020
44. Heterogeneity within Stratified Epithelial Stem Cell Populations Maintains the Oral Mucosa in Response to Physiological Stress
- Author
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Byrd, Kevin M, Piehl, Natalie C, Patel, Jeet H, Huh, Won Jae, Sequeira, Inês, Lough, Kendall J, Wagner, Bethany L, Marangoni, Pauline, Watt, Fiona M, Klein, Ophir D, Coffey, Robert J, and Williams, Scott E
- Subjects
Medical Biotechnology ,Biomedical and Clinical Sciences ,Stem Cell Research - Nonembryonic - Non-Human ,Regenerative Medicine ,Dental/Oral and Craniofacial Disease ,Stem Cell Research ,Underpinning research ,1.1 Normal biological development and functioning ,Animals ,Cell Division ,Cell Lineage ,Cells ,Cultured ,Female ,Flow Cytometry ,Fluorescence ,Immunohistochemistry ,Male ,Membrane Glycoproteins ,Mice ,Mouth Mucosa ,Nerve Tissue Proteins ,Stem Cells ,Wound Healing ,Igfbp5 ,Lrig1 ,label retention ,lineage tracing ,oral epithelium ,oriented cell division ,palate ,soft diet ,stem cell ,wound healing ,Biological Sciences ,Medical and Health Sciences ,Developmental Biology ,Biological sciences ,Biomedical and clinical sciences - Abstract
Stem cells in stratified epithelia are generally believed to adhere to a non-hierarchical single-progenitor model. Using lineage tracing and genetic label-retention assays, we show that the hard palatal epithelium of the oral cavity is unique in displaying marked proliferative heterogeneity. We identify a previously uncharacterized, infrequently-dividing stem cell population that resides within a candidate niche, the junctional zone (JZ). JZ stem cells tend to self-renew by planar symmetric divisions, respond to masticatory stresses, and promote wound healing, whereas frequently-dividing cells reside outside the JZ, preferentially renew through perpendicular asymmetric divisions, and are less responsive to injury. LRIG1 is enriched in the infrequently-dividing population in homeostasis, dynamically changes expression in response to tissue stresses, and promotes quiescence, whereas Igfbp5 preferentially labels a rapidly-growing, differentiation-prone population. These studies establish the oral mucosa as an important model system to study epithelial stem cell populations and how they respond to tissue stresses.
- Published
- 2019
45. A large pool of actively cycling progenitors orchestrates self-renewal and injury repair of an ectodermal appendage
- Author
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Sharir, Amnon, Marangoni, Pauline, Zilionis, Rapolas, Wan, Mian, Wald, Tomas, Hu, Jimmy K, Kawaguchi, Kyogo, Castillo-Azofeifa, David, Epstein, Leo, Harrington, Kyle, Pagella, Pierfrancesco, Mitsiadis, Thimios, Siebel, Christian W, Klein, Allon M, and Klein, Ophir D
- Subjects
Biological Sciences ,Genetics ,Stem Cell Research - Nonembryonic - Non-Human ,Regenerative Medicine ,Stem Cell Research ,1.1 Normal biological development and functioning ,Underpinning research ,Ameloblasts ,Animals ,Cell Differentiation ,Cell Division ,Cell Proliferation ,Ectoderm ,Epithelial Cells ,Incisor ,Mice ,Transgenic ,Signal Transduction ,Stem Cells ,Medical and Health Sciences ,Developmental Biology ,Biochemistry and cell biology - Abstract
The classical model of tissue renewal posits that small numbers of quiescent stem cells (SCs) give rise to proliferating transit-amplifying cells before terminal differentiation. However, many organs house pools of SCs with proliferative and differentiation potentials that diverge from this template. Resolving SC identity and organization is therefore central to understanding tissue renewal. Here, using a combination of single-cell RNA sequencing (scRNA-seq), mouse genetics and tissue injury approaches, we uncover cellular hierarchies and mechanisms that underlie the maintenance and repair of the continuously growing mouse incisor. Our results reveal that, during homeostasis, a group of actively cycling epithelial progenitors generates enamel-producing ameloblasts and adjacent layers of non-ameloblast cells. After injury, tissue repair was achieved through transient increases in progenitor-cell proliferation and through direct conversion of Notch1-expressing cells to ameloblasts. We elucidate epithelial SC identity, position and function, providing a mechanistic basis for the homeostasis and repair of a fast-turnover ectodermal appendage.
- Published
- 2019
46. Detecting 3D syndromic faces as outliers using unsupervised normalizing flow models
- Author
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Bannister, Jordan J., Wilms, Matthias, Aponte, J. David, Katz, David C., Klein, Ophir D., Bernier, Francois P.J., Spritz, Richard A., Hallgrímsson, Benedikt, and Forkert, Nils D.
- Published
- 2022
- Full Text
- View/download PDF
47. Perivascular NOTCH3+ stem cells drive meningioma tumorigenesis and resistance to radiotherapy
- Author
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Choudhury, Abrar, primary, Cady, Martha A., additional, Lucas, Calixto-Hope G., additional, Najem, Hinda, additional, Phillips, Joanna J., additional, Palikuqi, Brisa, additional, Zakimi, Naomi, additional, Joseph, Tara, additional, Birrueta, Janeth Ochoa., additional, Chen, William C., additional, Oberheim Bush, Nancy Ann, additional, Hervey-Jumper, Shawn L., additional, Klein, Ophir D., additional, Toedebusch, Christine M., additional, Horbinski, Craig M., additional, Magill, Stephen T., additional, Bhaduri, Aparna, additional, Perry, Arie, additional, Dickinson, Peter J., additional, Heimberger, Amy B., additional, Ashworth, Alan, additional, Crouch, Elizabeth E., additional, and Raleigh, David R., additional
- Published
- 2024
- Full Text
- View/download PDF
48. Downregulation of FGF Signaling by Spry4 Overexpression Leads to Shape Impairment, Enamel Irregularities, and Delayed Signaling Center Formation in the Mouse Molar
- Author
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Marangoni, Pauline, Charles, Cyril, Ahn, Youngwook, Seidel, Kerstin, Jheon, Andrew, Ganss, Bernhard, Krumlauf, Robb, Viriot, Laurent, and Klein, Ophir D
- Subjects
Biomedical and Clinical Sciences ,Dentistry ,Dental/Oral and Craniofacial Disease ,Genetics ,Aetiology ,2.1 Biological and endogenous factors ,FGF SIGNALING ,ENAMEL MINERALIZATION DEFECT ,TOOTH DEVELOPMENT ,SPRY4 ,Biomedical and clinical sciences - Abstract
FGF signaling plays a critical role in tooth development, and mutations in modulators of this pathway produce a number of striking phenotypes. However, many aspects of the role of the FGF pathway in regulating the morphological features and the mineral quality of the dentition remain unknown. Here, we used transgenic mice overexpressing the FGF negative feedback regulator Sprouty4 under the epithelial keratin 14 promoter (K14-Spry4) to achieve downregulation of signaling in the epithelium. This led to highly penetrant defects affecting both cusp morphology and the enamel layer. We characterized the phenotype of erupted molars, identified a developmental delay in K14-Spry4 transgenic embryos, and linked this with changes in the tooth developmental sequence. These data further delineate the role of FGF signaling in the development of the dentition and implicate the pathway in the regulation of tooth mineralization. © 2019 The Authors. JBMR Plus is published by Wiley Periodicals, Inc. on behalf of American Society for Bone and Mineral Research.
- Published
- 2019
49. Sonic Hedgehog Signaling Is Required for Cyp26 Expression during Embryonic Development.
- Author
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El Shahawy, Maha, Reibring, Claes-Göran, Hallberg, Kristina, Neben, Cynthia L, Marangoni, Pauline, Harfe, Brian D, Klein, Ophir D, Linde, Anders, and Gritli-Linde, Amel
- Subjects
Epithelial Cells ,Tooth ,Animals ,Mice ,Tretinoin ,Signal Transduction ,Apoptosis ,Cell Differentiation ,Gene Expression Regulation ,Developmental ,Embryonic Development ,Organogenesis ,Hedgehog Proteins ,Embryo ,Mammalian ,Retinoic Acid 4-Hydroxylase ,Cytochrome P450 Family 26 ,CRE/LoxP ,Cyp26 enzymes ,congenital anomalies ,hedgehog signaling ,mouse models ,retinoic acid ,smoothened ,sonic hedgehog ,CRE ,LoxP ,Gene Expression Regulation ,Developmental ,Embryo ,Mammalian ,Other Chemical Sciences ,Genetics ,Other Biological Sciences ,Chemical Physics - Abstract
Deciphering how signaling pathways interact during development is necessary for understanding the etiopathogenesis of congenital malformations and disease. In several embryonic structures, components of the Hedgehog and retinoic acid pathways, two potent players in development and disease are expressed and operate in the same or adjacent tissues and cells. Yet whether and, if so, how these pathways interact during organogenesis is, to a large extent, unclear. Using genetic and experimental approaches in the mouse, we show that during development of ontogenetically different organs, including the tail, genital tubercle, and secondary palate, Sonic hedgehog (SHH) loss-of-function causes anomalies phenocopying those induced by enhanced retinoic acid signaling and that SHH is required to prevent supraphysiological activation of retinoic signaling through maintenance and reinforcement of expression of the Cyp26 genes. Furthermore, in other tissues and organs, disruptions of the Hedgehog or the retinoic acid pathways during development generate similar phenotypes. These findings reveal that rigidly calibrated Hedgehog and retinoic acid activities are required for normal organogenesis and tissue patterning.
- Published
- 2019
50. Feedback regulation of RTK signaling in development
- Author
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Neben, Cynthia L, Lo, Megan, Jura, Natalia, and Klein, Ophir D
- Subjects
Biochemistry and Cell Biology ,Biomedical and Clinical Sciences ,Biological Sciences ,Genetics ,Underpinning research ,2.1 Biological and endogenous factors ,1.1 Normal biological development and functioning ,Aetiology ,Generic health relevance ,Animals ,Embryo ,Mammalian ,Embryonic Development ,Gene Expression Regulation ,Developmental ,Humans ,Neoplasm Proteins ,Neoplasms ,Receptor Protein-Tyrosine Kinases ,Signal Transduction ,Receptor tyrosine kinases ,Signaling pathways ,Feedback regulation ,Developmental control ,RASopathies ,Sprouty ,Medical and Health Sciences ,Developmental Biology ,Biological sciences ,Biomedical and clinical sciences ,Health sciences - Abstract
Precise regulation of the amplitude and duration of receptor tyrosine kinase (RTK) signaling is critical for the execution of cellular programs and behaviors. Understanding these control mechanisms has important implications for the field of developmental biology, and in recent years, the question of how augmentation or attenuation of RTK signaling via feedback loops modulates development has become of increasing interest. RTK feedback regulation is also important for human disease research; for example, germline mutations in genes that encode RTK signaling pathway components cause numerous human congenital syndromes, and somatic alterations contribute to the pathogenesis of diseases such as cancers. In this review, we survey regulators of RTK signaling that tune receptor activity and intracellular transduction cascades, with a focus on the roles of these genes in the developing embryo. We detail the diverse inhibitory mechanisms utilized by negative feedback regulators that, when lost or perturbed, lead to aberrant increases in RTK signaling. We also discuss recent biochemical and genetic insights into positive regulators of RTK signaling and how these proteins function in tandem with negative regulators to guide embryonic development.
- Published
- 2019
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