Matteo Amitaba Mole'

All Publications

• A single cell characterisation of human embryogenesis identifies pluripotency transitions and putative anterior hypoblast centre NATURE COMMUNICATIONS Mole, M. A., Coorens, T. H., Shahbazi, M. N., Weberling, A., Weatherbee, B. T., Gantner, C. W., Sancho-Serra, C., Richardson, L., Drinkwater, A., Syed, N., Engley, S., Snell, P., Christie, L., Elder, K., Campbell, A., Fishel, S., Behjati, S., Vento-Tormo, R., Zernicka-Goetz, M. 2021; 12 (1): 3679

  Abstract

  Following implantation, the human embryo undergoes major morphogenetic transformations that establish the future body plan. While the molecular events underpinning this process are established in mice, they remain unknown in humans. Here we characterise key events of human embryo morphogenesis, in the period between implantation and gastrulation, using single-cell analyses and functional studies. First, the embryonic epiblast cells transition through different pluripotent states and act as a source of FGF signals that ensure proliferation of both embryonic and extra-embryonic tissues. In a subset of embryos, we identify a group of asymmetrically positioned extra-embryonic hypoblast cells expressing inhibitors of BMP, NODAL and WNT signalling pathways. We suggest that this group of cells can act as the anterior singalling centre to pattern the epiblast. These results provide insights into pluripotency state transitions, the role of FGF signalling and the specification of anterior-posterior axis during human embryo development.

  View details for DOI 10.1038/s41467-021-23758-w

  View details for Web of Science ID 000665032700001

  View details for PubMedID 34140473

  View details for PubMedCentralID PMC8211662

• Integrin beta 1 coordinates survival and morphogenesis of the embryonic lineage upon implantation and pluripotency transition CELL REPORTS Mole, M., Weberling, A., Faessler, R., Campbell, A., Fishel, S., Zernicka-Goetz, M. 2021; 34 (10): 108834

  Abstract

  At implantation, the embryo establishes contacts with the maternal endometrium. This stage is associated with a high incidence of preclinical pregnancy losses. While the maternal factors underlying uterine receptivity have been investigated, the signals required by the embryo for successful peri-implantation development remain elusive. To explore these, we studied integrin β1 signaling, as embryos deficient for this receptor degenerate at implantation. We demonstrate that the coordinated action of pro-survival signals and localized actomyosin suppression via integrin β1 permits the development of the embryo beyond implantation. Failure of either process leads to developmental arrest and apoptosis. Pharmacological stimulation through fibroblast growth factor 2 (FGF2) and insulin-like growth factor 1 (IGF1), coupled with ROCK-mediated actomyosin inhibition, rescues the deficiency of integrin β1, promoting progression to post-implantation stages. Mutual exclusion between integrin β1 and actomyosin seems to be conserved in the human embryo, suggesting the possibility that these mechanisms could also underlie the transition of the human epiblast from pre- to post-implantation.

  View details for DOI 10.1016/j.celrep.2021.108834

  View details for Web of Science ID 000627660900021

  View details for PubMedID 33691117

  View details for PubMedCentralID PMC7966855

• Integrin-Mediated Focal Anchorage Drives Epithelial Zippering during Mouse Neural Tube Closure DEVELOPMENTAL CELL Mole, M. A., Galea, G. L., Rolo, A., Weberling, A., Nychyk, O., De Castro, S. C., Savery, D., Faessler, R., Ybot-Gonzalez, P., Greene, N. E., Copp, A. J. 2020; 52 (3): 321-+

  Abstract

  Epithelial fusion is a key process of morphogenesis by which tissue connectivity is established between adjacent epithelial sheets. A striking and poorly understood feature of this process is "zippering," whereby a fusion point moves directionally along an organ rudiment. Here, we uncover the molecular mechanism underlying zippering during mouse spinal neural tube closure. Fusion is initiated via local activation of integrin β1 and focal anchorage of surface ectoderm cells to a shared point of fibronectin-rich basement membrane, where the neural folds first contact each other. Surface ectoderm cells undergo proximal junction shortening, establishing a transitory semi-rosette-like structure at the zippering point that promotes juxtaposition of cells across the midline enabling fusion propagation. Tissue-specific ablation of integrin β1 abolishes the semi-rosette formation, preventing zippering and causing spina bifida. We propose integrin-mediated anchorage as an evolutionarily conserved mechanism of general relevance for zippering closure of epithelial gaps whose disturbance can produce clinically important birth defects.

  View details for DOI 10.1016/j.devcel.2020.01.012

  View details for Web of Science ID 000513784400010

  View details for PubMedID 32049039

  View details for PubMedCentralID PMC7008250

• Comparative analysis of human and mouse development: From zygote to pre-gastrulation GASTRULATION: FROM EMBRYONIC PATTERN TO FORM Mole, M. A., Weberling, A., Zernicka-Goetz, M., SolnicaKrezel, L. 2020; 136: 113-+

  Abstract

  Development of the mammalian embryo begins with formation of the totipotent zygote during fertilization. This initial cell is able to give rise to every embryonic tissue of the developing organism as well as all extra-embryonic lineages, such as the placenta and the yolk sac, which are essential for the initial patterning and support growth of the fetus until birth. As the embryo transits from pre- to post-implantation, major structural and transcriptional changes occur within the embryonic lineage to set up the basis for the subsequent phase of gastrulation. Fine-tuned coordination of cell division, morphogenesis and differentiation is essential to ultimately promote assembly of the future fetus. Here, we review the current knowledge of mammalian development of both mouse and human focusing on morphogenetic processes leading to the onset of gastrulation, when the embryonic anterior-posterior axis becomes established and the three germ layers start to be specified.

  View details for DOI 10.1016/bs.ctdb.2019.10.002

  View details for Web of Science ID 000611830600005

  View details for PubMedID 31959285

• Cellular basis of neuroepithelial bending during mouse spinal neural tube closure DEVELOPMENTAL BIOLOGY McShane, S. G., Mole, M. A., Savery, D., Greene, N. E., Tam, P. L., Copp, A. J. 2015; 404 (2): 113-124

  Abstract

  Bending of the neural plate at paired dorsolateral hinge points (DLHPs) is required for neural tube closure in the spinal region of the mouse embryo. As a step towards understanding the morphogenetic mechanism of DLHP development, we examined variations in neural plate cellular architecture and proliferation during closure. Neuroepithelial cells within the median hinge point (MHP) contain nuclei that are mainly basally located and undergo relatively slow proliferation, with a 7 h cell cycle length. In contrast, cells in the dorsolateral neuroepithelium, including the DLHP, exhibit nuclei distributed throughout the apico-basal axis and undergo rapid proliferation, with a 4 h cell cycle length. As the neural folds elevate, cell numbers increase to a greater extent in the dorsolateral neural plate that contacts the surface ectoderm, compared with the more ventromedial neural plate where cells contact paraxial mesoderm and notochord. This marked increase in dorsolateral cell number cannot be accounted for solely on the basis of enhanced cell proliferation in this region. We hypothesised that neuroepithelial cells may translocate in a ventral-to-dorsal direction as DLHP formation occurs, and this was confirmed by vital cell labelling in cultured embryos. The translocation of cells into the neural fold, together with its more rapid cell proliferation, leads to an increase in cell density dorsolaterally compared with the more ventromedial neural plate. These findings suggest a model in which DLHP formation may proceed through 'buckling' of the neuroepithelium at a dorso-ventral boundary marked by a change in cell-packing density.

  View details for DOI 10.1016/j.ydbio.2015.06.003

  View details for Web of Science ID 000358815700010

  View details for PubMedID 26079577

  View details for PubMedCentralID PMC4528075

• Live-Imaging Analysis of Epithelial Zippering During Mouse Neural Tube Closure. Methods in molecular biology (Clifton, N.J.) Mole, M. A., Galea, G. L., Copp, A. J. 2023; 2608: 147-162

  Abstract

  Zippering is a phenomenon of tissue morphogenesis whereby fusion between opposing epithelia progresses unidirectionally over significant distances, similar to the travel of a zip fastener, to ultimately ensure closure of an opening. A comparable process can be observed during Drosophila dorsal closure and mammalian wound healing, while zippering is employed by numerous organs such as the optic fissure, palatal shelves, tracheoesophageal foregut, and presumptive genitalia to mediate tissue sealing during normal embryonic development. Particularly striking is zippering propagation during neural tube morphogenesis, where the fusion point travels extensively along the embryonic axis to ensure closure of the neural tube. Advances in time-lapse microscopy and culture conditions have opened the opportunity for successful imaging of whole-mouse embryo development over time, providing insights into the precise cellular behavior underlying zippering propagation. Studies in mouse and the ascidian Ciona have revealed the fine-tuned cell shape changes and junction remodeling which occur at the site of zippering during neural tube morphogenesis. Here, we describe a step-by-step method for imaging at single-cell resolution the process of zippering and tissue remodeling which occurs during closure of the spinal neural tube in mouse. We also provide instructions and suggestions for quantitative morphometric analysis of cell behavior during zippering progression. This procedure can be further combined with genetic mutant models (e.g., knockouts), offering the possibility of studying the dynamics of tissue fusion and zippering propagation, which underlie a wide range of open neural tube defects.

  View details for DOI 10.1007/978-1-0716-2887-4_10

  View details for PubMedID 36653707

• Vangl2-environment interaction causes severe neural tube defects, without abnormal neuroepithelial convergent extension DISEASE MODELS & MECHANISMS Nychyk, O., Galea, G. L., Mole, M., Savery, D., Greene, N. E., Stanier, P., Copp, A. J. 2022; 15 (1)

  Abstract

  Planar cell polarity (PCP) signalling is vital for initiation of mouse neurulation, with diminished convergent extension (CE) cell movements leading to craniorachischisis, a severe neural tube defect (NTD). Some humans with NTDs also have PCP gene mutations but these are heterozygous, not homozygous as in mice. Other genetic or environmental factors may interact with partial loss of PCP function in human NTDs. We found that reduced sulfation of glycosaminoglycans interacts with heterozygosity for the Lp allele of Vangl2 (a core PCP gene), to cause craniorachischisis in cultured mouse embryos, with rescue by exogenous sulphate. We hypothesized that this glycosaminoglycan-PCP interaction may regulate CE, but, surprisingly, DiO labelling of the embryonic node demonstrates no abnormality of midline axial extension in sulfation-depleted Lp/+ embryos. Positive-control Lp/Lp embryos show severe CE defects. Abnormalities were detected in the size and shape of somites that flank the closing neural tube in sulfation-depleted Lp/+ embryos. We conclude that failure of closure initiation can arise by a mechanism other than faulty neuroepithelial CE, with possible involvement of matrix-mediated somite expansion, adjacent to the closing neural tube.

  View details for DOI 10.1242/dmm.049194

  View details for Web of Science ID 000751658700002

  View details for PubMedID 34842271

  View details for PubMedCentralID PMC8807581

• The role of cell-ECM adhesions in mouse neural tube closure Papastergios, E. V., Mole, M. A., Galea, G. L., Greene, N. E., Copp, A. J. WILEY. 2021: 959

  View details for Web of Science ID 000697117200084

• Human embryo polarization requires PLC signaling to mediate trophectoderm specification ELIFE Zhu, M., Shahbazi, M., Martin, A., Zhang, C., Sozen, B., Borsos, M., Mandelbaum, R. S., Paulson, R. J., Mole, M. A., Esbert, M., Titus, S., Scott, R. T., Campbell, A., Fishel, S., Gradinaru, V., Zhao, H., Wu, K., Chen, Z., Seli, E., de los Santos, M. J., Zernicka Goetz, M. 2021; 10

  Abstract

  Apico-basal polarization of cells within the embryo is critical for the segregation of distinct lineages during mammalian development. Polarized cells become the trophectoderm (TE), which forms the placenta, and apolar cells become the inner cell mass (ICM), the founding population of the fetus. The cellular and molecular mechanisms leading to polarization of the human embryo and its timing during embryogenesis have remained unknown. Here, we show that human embryo polarization occurs in two steps: it begins with the apical enrichment of F-actin and is followed by the apical accumulation of the PAR complex. This two-step polarization process leads to the formation of an apical domain at the 8-16 cell stage. Using RNA interference, we show that apical domain formation requires Phospholipase C (PLC) signaling, specifically the enzymes PLCB1 and PLCE1, from the eight-cell stage onwards. Finally, we show that although expression of the critical TE differentiation marker GATA3 can be initiated independently of embryo polarization, downregulation of PLCB1 and PLCE1 decreases GATA3 expression through a reduction in the number of polarized cells. Therefore, apical domain formation reinforces a TE fate. The results we present here demonstrate how polarization is triggered to regulate the first lineage segregation in human embryos.

  View details for DOI 10.7554/eLife.65068.sa2

  View details for Web of Science ID 000709351100001

  View details for PubMedID 34569938

  View details for PubMedCentralID PMC8514238

• Modelling the impact of decidual senescence on embryo implantation in human endometrial assembloids ELIFE Rawlings, T. M., Makwana, K., Taylor, D. M., Mole, M. A., Fishwick, K. J., Tryfonos, M., Odendaal, J., Hawkes, A., Zernicka-Goetz, M., Hartshorne, G. M., Brosens, J. J., Lucas, E. S. 2021; 10

  Abstract

  Decidual remodelling of midluteal endometrium leads to a short implantation window after which the uterine mucosa either breaks down or is transformed into a robust matrix that accommodates the placenta throughout pregnancy. To gain insights into the underlying mechanisms, we established and characterized endometrial assembloids, consisting of gland-like organoids and primary stromal cells. Single-cell transcriptomics revealed that decidualized assembloids closely resemble midluteal endometrium, harbouring differentiated and senescent subpopulations in both glands and stroma. We show that acute senescence in glandular epithelium drives secretion of multiple canonical implantation factors, whereas in the stroma it calibrates the emergence of anti-inflammatory decidual cells and pro-inflammatory senescent decidual cells. Pharmacological inhibition of stress responses in pre-decidual cells accelerated decidualization by eliminating the emergence of senescent decidual cells. In co-culture experiments, accelerated decidualization resulted in entrapment of collapsed human blastocysts in a robust, static decidual matrix. By contrast, the presence of senescent decidual cells created a dynamic implantation environment, enabling embryo expansion and attachment, although their persistence led to gradual disintegration of assembloids. Our findings suggest that decidual senescence controls endometrial fate decisions at implantation and highlight how endometrial assembloids may accelerate the discovery of new treatments to prevent reproductive failure.

  View details for DOI 10.7554/eLife.69603.sa2

  View details for Web of Science ID 000709339300001

  View details for PubMedID 34487490

  View details for PubMedCentralID PMC8523170

• The role of Sox2 in neuromesodermal progenitors and neural specification in the mouse Papastergios, E. V., Mugele, D., Moulding, D. A., Savery, D., Mole, M. A., Greene, N. E., Martinez-Barbera, J., Copp, A. J. WILEY. 2020: 338

  View details for Web of Science ID 000524885900615

• Vangl2 disruption alters the biomechanics of late spinal neurulation leading to spina bifida in mouse embryos DISEASE MODELS & MECHANISMS Galea, G. L., Nychyk, O., Mole, M. A., Moulding, D., Savery, D., Nikolopoulou, E., Henderson, D. J., Greene, N. E., Copp, A. J. 2018; 11 (3)

  Abstract

  Human mutations in the planar cell polarity component VANGL2 are associated with the neural tube defect spina bifida. Homozygous Vangl2 mutation in mice prevents initiation of neural tube closure, precluding analysis of its subsequent roles in neurulation. Spinal neurulation involves rostral-to-caudal 'zippering' until completion of closure is imminent, when a caudal-to-rostral closure point, 'Closure 5', arises at the caudal-most extremity of the posterior neuropore (PNP). Here, we used Grhl3Cre to delete Vangl2 in the surface ectoderm (SE) throughout neurulation and in an increasing proportion of PNP neuroepithelial cells at late neurulation stages. This deletion impaired PNP closure after the ∼25-somite stage and resulted in caudal spina bifida in 67% of Grhl3Cre/+Vangl2Fl/Fl embryos. In the dorsal SE, Vangl2 deletion diminished rostrocaudal cell body orientation, but not directional polarisation of cell divisions. In the PNP, Vangl2 disruption diminished mediolateral polarisation of apical neuroepithelial F-actin profiles and resulted in eversion of the caudal PNP. This eversion prevented elevation of the caudal PNP neural folds, which in control embryos is associated with formation of Closure 5 around the 25-somite stage. Closure 5 formation in control embryos is associated with a reduction in mechanical stress withstood at the main zippering point, as inferred from the magnitude of neural fold separation following zippering point laser ablation. This stress accommodation did not happen in Vangl2-disrupted embryos. Thus, disruption of Vangl2-dependent planar-polarised processes in the PNP neuroepithelium and SE preclude zippering point biomechanical accommodation associated with Closure 5 formation at the completion of PNP closure.

  View details for DOI 10.1242/dmm.032219

  View details for Web of Science ID 000429382700006

  View details for PubMedID 29590636

  View details for PubMedCentralID PMC5897727

• Biomechanical coupling of the closing spinal neural tube facilitates neural fold apposition Galea, G. L., Cho, Y., Galea, G., Mole, M. A., Rolo, A., Savery, D., Moulding, D., Nikolopoulou, E., Greene, N. E., Copp, A. J. HINDAWI LTD. 2017

  View details for Web of Science ID 000405659000014

• Biomechanical coupling facilitates spinal neural tube closure in mouse embryos PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Galea, G. L., Cho, Y., Galea, G., Mole, M. A., Rolo, A., Savery, D., Moulding, D., Culshaw, L. H., Nikolopoulou, E., Greene, N. E., Copp, A. J. 2017; 114 (26): E5177-E5186

  Abstract

  Neural tube (NT) formation in the spinal region of the mammalian embryo involves a wave of "zippering" that passes down the elongating spinal axis, uniting the neural fold tips in the dorsal midline. Failure of this closure process leads to open spina bifida, a common cause of severe neurologic disability in humans. Here, we combined a tissue-level strain-mapping workflow with laser ablation of live-imaged mouse embryos to investigate the biomechanics of mammalian spinal closure. Ablation of the zippering point at the embryonic dorsal midline causes far-reaching, rapid separation of the elevating neural folds. Strain analysis revealed tissue expansion around the zippering point after ablation, but predominant tissue constriction in the caudal and ventral neural plate zone. This zone is biomechanically coupled to the zippering point by a supracellular F-actin network, which includes an actin cable running along the neural fold tips. Pharmacologic inhibition of F-actin or laser ablation of the cable causes neural fold separation. At the most advanced somite stages, when completion of spinal closure is imminent, the cable forms a continuous ring around the neuropore, and simultaneously, a new caudal-to-rostral zippering point arises. Laser ablation of this new closure initiation point causes neural fold separation, demonstrating its biomechanical activity. Failure of spinal closure in pre-spina bifida Zic2Ku mutant embryos is associated with altered tissue biomechanics, as indicated by greater neuropore widening after ablation. Thus, this study identifies biomechanical coupling of the entire region of active spinal neurulation in the mouse embryo as a prerequisite for successful NT closure.

  View details for DOI 10.1073/pnas.1700934114

  View details for Web of Science ID 000404108400021

  View details for PubMedID 28607062

  View details for PubMedCentralID PMC5495245

• Regulation of cell protrusions by small GTPases during fusion of the neural folds ELIFE Rolo, A., Savery, D., Escuin, S., de Castro, S. C., Armer, H. J., Munro, P. G., Mole, M. A., Greene, N. E., Copp, A. J. 2016; 5: e13273

  Abstract

  Epithelial fusion is a crucial process in embryonic development, and its failure underlies several clinically important birth defects. For example, failure of neural fold fusion during neurulation leads to open neural tube defects including spina bifida. Using mouse embryos, we show that cell protrusions emanating from the apposed neural fold tips, at the interface between the neuroepithelium and the surface ectoderm, are required for completion of neural tube closure. By genetically ablating the cytoskeletal regulators Rac1 or Cdc42 in the dorsal neuroepithelium, or in the surface ectoderm, we show that these protrusions originate from surface ectodermal cells and that Rac1 is necessary for the formation of membrane ruffles which typify late closure stages, whereas Cdc42 is required for the predominance of filopodia in early neurulation. This study provides evidence for the essential role and molecular regulation of membrane protrusions prior to fusion of a key organ primordium in mammalian development.

  View details for DOI 10.7554/elife.13273

  View details for Web of Science ID 000374827600001

  View details for PubMedID 27114066

  View details for PubMedCentralID PMC4846376

• Cell-matrix interactions and cell dynamics of neuroepithelial bending during mouse spinal neural tube closure Mole, M. A., Mugele, D., Ybot-Gonzalez, P., Greene, N. E., Copp, A. J. HINDAWI LTD. 2015

  View details for Web of Science ID 000367181100029