Madeline Hughes Cooper

All Publications

• Nanobodies against the myelin enzyme CNPase as tools for structural and functional studies. bioRxiv : the preprint server for biology Markusson, S., Raasakka, A., Schröder, M., Sograte-Idrissi, S., Rahimi, A. M., Asadpour, O., Körner, H., Lodygin, D., Eichel-Vogel, M. A., Chowdhury, R., Sutinen, A., Muruganandam, G., Iyer, M., Cooper, M. H., Weigel, M. K., Ambiel, N., Werner, H. B., Zuchero, J. B., Opazo, F., Kursula, P. 2024

  Abstract

  2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) is an abundant constituent of central nervous system non-compact myelin, frequently used as a marker antigen for myelinating cells. The catalytic activity of CNPase, the 3'-hydrolysis of 2',3'-cyclic nucleotides, is well characterised in vitro, but the in vivo function of CNPase remains unclear. CNPase interacts with the actin cytoskeleton to counteract the developmental closure of cytoplasmic channels that travel through compact myelin; its enzymatic activity may be involved in adenosine metabolism and RNA degradation. We developed a set of high-affinity nanobodies recognizing the phosphodiesterase domain of CNPase, and the crystal structures of each complex show that the five nanobodies have distinct epitopes. One of the nanobodies bound deep into the CNPase active site and acted as an inhibitor. Moreover, the nanobodies were characterised in imaging applications and as intrabodies, expressed in mammalian cells, such as primary oligodendrocytes. Fluorescently labelled nanobodies functioned in imaging of teased nerve fibers and whole brain tissue sections, as well as super-resolution microscopy. These anti-CNPase nanobodies provide new tools for structural and functional biology of myelination, including high-resolution imaging of nerve tissue.

  View details for DOI 10.1101/2024.05.25.595513

  View details for PubMedID 38826303

  View details for PubMedCentralID PMC11142274

• Optical Control of G-Actin with a Photoswitchable Latrunculin. Journal of the American Chemical Society Vepřek, N. A., Cooper, M. H., Laprell, L., Yang, E. J., Folkerts, S., Bao, R., Boczkowska, M., Palmer, N. J., Dominguez, R., Oertner, T. G., Pon, L. A., Zuchero, J. B., Trauner, D. H. 2024

  Abstract

  Actin is one of the most abundant proteins in eukaryotic cells and is a key component of the cytoskeleton. A range of small molecules has emerged that interfere with actin dynamics by either binding to polymeric F-actin or monomeric G-actin to stabilize or destabilize filaments or prevent their formation and growth, respectively. Among these, the latrunculins, which bind to G-actin and affect polymerization, are widely used as tools to investigate actin-dependent cellular processes. Here, we report a photoswitchable version of latrunculin, termed opto-latrunculin (OptoLat), which binds to G-actin in a light-dependent fashion and affords optical control over actin polymerization. OptoLat can be activated with 390-490 nm pulsed light and rapidly relaxes to its inactive form in the dark. Light activated OptoLat induced depolymerization of F-actin networks in oligodendrocytes and budding yeast, as shown by fluorescence microscopy. Subcellular control of actin dynamics in human cancer cell lines was demonstrated via live cell imaging. Light-activated OptoLat also reduced microglia surveillance in organotypic mouse brain slices while ramification was not affected. Incubation in the dark did not alter the structural and functional integrity of the microglia. Together, our data demonstrate that OptoLat is a useful tool for the elucidation of G-actin dependent dynamic processes in cells and tissues.

  View details for DOI 10.1021/jacs.3c10776

  View details for PubMedID 38511265

• Oligodendrocyte calcium signaling promotes actin-dependent myelin sheath extension. Nature communications Iyer, M., Kantarci, H., Cooper, M. H., Ambiel, N., Novak, S. W., Andrade, L. R., Lam, M., Jones, G., Münch, A. E., Yu, X., Khakh, B. S., Manor, U., Zuchero, J. B. 2024; 15 (1): 265

  Abstract

  Myelin is essential for rapid nerve signaling and is increasingly found to play important roles in learning and in diverse diseases of the CNS. Morphological parameters of myelin such as sheath length are thought to precisely tune conduction velocity, but the mechanisms controlling sheath morphology are poorly understood. Local calcium signaling has been observed in nascent myelin sheaths and can be modulated by neuronal activity. However, the role of calcium signaling in sheath formation remains incompletely understood. Here, we use genetic tools to attenuate oligodendrocyte calcium signaling during myelination in the developing mouse CNS. Surprisingly, genetic calcium attenuation does not grossly affect the number of myelinated axons or myelin thickness. Instead, calcium attenuation causes myelination defects resulting in shorter, dysmorphic sheaths. Mechanistically, calcium attenuation reduces actin filaments in oligodendrocytes, and an intact actin cytoskeleton is necessary and sufficient to achieve accurate myelin morphology. Together, our work reveals a cellular mechanism required for accurate CNS myelin formation and may provide mechanistic insight into how oligodendrocytes respond to neuronal activity to sculpt and refine myelin sheaths.

  View details for DOI 10.1038/s41467-023-44238-3

  View details for PubMedID 38177161

  View details for PubMedCentralID 9651929

• CNS myelination requires VAMP2/3-mediated membrane expansion in oligodendrocytes. Nature communications Lam, M., Takeo, K., Almeida, R. G., Cooper, M. H., Wu, K., Iyer, M., Kantarci, H., Zuchero, J. B. 2022; 13 (1): 5583

  Abstract

  Myelin is required for rapid nerve signaling and is emerging as a key driver of CNS plasticity and disease. How myelin is built and remodeled remains a fundamental question of neurobiology. Central to myelination is the ability of oligodendrocytes to add vast amounts of new cell membrane, expanding their surface areas by many thousand-fold. However, how oligodendrocytes add new membrane to build or remodel myelin is not fully understood. Here, we show that CNS myelin membrane addition requires exocytosis mediated by the vesicular SNARE proteins VAMP2/3. Genetic inactivation of VAMP2/3 in myelinating oligodendrocytes caused severe hypomyelination and premature death without overt loss of oligodendrocytes. Through live imaging, we discovered that VAMP2/3-mediated exocytosis drives membrane expansion within myelin sheaths to initiate wrapping and power sheath elongation. In conjunction with membrane expansion, mass spectrometry of oligodendrocyte surface proteins revealed that VAMP2/3 incorporates axon-myelin adhesion proteins that are collectively required to form nodes of Ranvier. Together, our results demonstrate that VAMP2/3-mediated membrane expansion in oligodendrocytes is indispensable for myelin formation, uncovering a cellular pathway that could sculpt myelination patterns in response to activity-dependent signals or be therapeutically targeted to promote regeneration in disease.

  View details for DOI 10.1038/s41467-022-33200-4

  View details for PubMedID 36151203

• Modeling myelin: A toolkit for exploring myelin's mysteries in vitro. Developmental cell Cooper, M. H., Zuchero, J. B. 2021; 56 (9): 1215–17

  Abstract

  In the myelin field, there is a lack of reliable in vitro tools to study myelination, especially using human cells. In this issue of Developmental Cell, James et al. present a guide to generating human iPSC-derived "myelinoids"-3D models of myelination that reliably achieve mature myelin formation.

  View details for DOI 10.1016/j.devcel.2021.04.015

  View details for PubMedID 33945781

• Soft extracellular matrix enhances inflammatory activation of mesenchymal stromal cells to induce monocyte production and trafficking SCIENCE ADVANCES Wong, S., Lenzini, S., Cooper, M. H., Mooney, D. J., Shin, J. 2020; 6 (15): eaaw0158

  Abstract

  Mesenchymal stromal cells (MSCs) modulate immune cells to ameliorate multiple inflammatory pathologies. Biophysical signals that regulate this process are poorly defined. By engineering hydrogels with tunable biophysical parameters relevant to bone marrow where MSCs naturally reside, we show that soft extracellular matrix maximizes the ability of MSCs to produce paracrine factors that have been implicated in monocyte production and chemotaxis upon inflammatory stimulation by tumor necrosis factor-α (TNFα). Soft matrix increases clustering of TNF receptors, thereby enhancing NF-κB activation and downstream gene expression. Actin polymerization and lipid rafts, but not myosin-II contractility, regulate mechanosensitive activation of MSCs by TNFα. We functionally demonstrate that human MSCs primed with TNFα in soft matrix enhance production of human monocytes in marrow of xenografted mice and increase trafficking of monocytes via CCL2. The results suggest the importance of biophysical signaling in tuning inflammatory activation of stromal cells to control the innate immune system.

  View details for DOI 10.1126/sciadv.aaw0158

  View details for Web of Science ID 000525751400002

  View details for PubMedID 32284989

  View details for PubMedCentralID PMC7141831

• Chapter 1 - Design principles for dynamic microphysiological systems Microfluidic Cell Culture Systems Cooper, M., Charest, J. L., Coppeta, J. Elsevier Inc.. 2019; 2: 1–29

  View details for DOI 10.1016/B978-0-12-813671-3.00001-3

• Community health workers on a college campus: Effects on influenza vaccination JOURNAL OF AMERICAN COLLEGE HEALTH Huang, J. J., Francesconi, M., Cooper, M. H., Covello, A., Guo, M., Gharib, S. D. 2018; 66 (4): 317–23

  Abstract

  To assess the impact of a campus community health worker program (HealthPALs) on student influenza vaccination.Undergraduate students at a northeastern US university (enrollment 6650), influenza seasons 2011-2012 through 2015-2016.Study design: Difference-in-differences analysis of student vaccination at campus dormitory influenza clinics during intervention vs. baseline.In the first intervention year, HealthPALs conducted in-person peer outreach at several campus dormitory flu clinics. Subsequent years, HealthPALs conducted an enhanced intervention, with the addition of a personalized, dormitory-specific social media campaign appealing to students' community identity.The initial intervention increased vaccinations by 66% (IRR = 1.66, 95%CI 1.39-1.97) at intervention clinics relative to control. The enhanced intervention increased vaccinations by 85% (IRR = 1.85, 95%CI 1.75-1.96).Community health workers can be a highly effective, low-cost strategy for increasing influenza vaccination among college students. This model could also be used to address other campus health challenges where student engagement is key.

  View details for PubMedID 29447623

• Deterministic encapsulation of single cells in thin tunable microgels for niche modelling and therapeutic delivery. Nature materials Mao, A. S., Shin, J., Utech, S., Wang, H., Uzun, O., Li, W., Cooper, M., Hu, Y., Zhang, L., Weitz, D. A., Mooney, D. J. 2017; 16 (2): 236-243

  Abstract

  Existing techniques to encapsulate cells into microscale hydrogels generally yield high polymer-to-cell ratios and lack control over the hydrogel's mechanical properties. Here, we report a microfluidic-based method for encapsulating single cells in an approximately six-micrometre layer of alginate that increases the proportion of cell-containing microgels by a factor of ten, with encapsulation efficiencies over 90%. We show that in vitro cell viability was maintained over a three-day period, that the microgels are mechanically tractable, and that, for microscale cell assemblages of encapsulated marrow stromal cells cultured in microwells, osteogenic differentiation of encapsulated cells depends on gel stiffness and cell density. We also show that intravenous injection of singly encapsulated marrow stromal cells into mice delays clearance kinetics and sustains donor-derived soluble factors in vivo. The encapsulation of single cells in tunable hydrogels should find use in a variety of tissue engineering and regenerative medicine applications.

  View details for DOI 10.1038/nmat4781

  View details for PubMedID 27798621

  View details for PubMedCentralID PMC5372217