Mammalian Cell Biology

Genetics, cell biology and development

Our research is developed in two main and complementary axes: the cell biology of proliferation and that of myogenic differentiation of mammalian cells.
Reversible protein phosphorylation is an essential component in the transduction of signals associated with normal and cancer cell proliferation as well as cell cycle arrest and exit into myogenic differentiation. Using cell biology approaches on primary and established mammalian cultured cells and adult stem cell isolated from skeletal muscle we have shown the implication of protein post-translational modifications such as phosphorylation in the signalling pathways regulating mammalian cell proliferation and differentiation. In a more recent development of our research we are examining the multi-lineage differentiation potential of adult muscle stem cells and their interest for regenerative medicine.

Cell Cycle Regulation and Cancer Signaling

We have identified key points in the crosstalk of major multi-tasking enzymes, such as cAMP-dependent Protein Kinase (PKA), Akt/PKB family kinases and phosphatase 2A (PP2A) in the modulation Cyclin-Dependent Kinases (CDK) during cell cycle progression. This crosstalk is the target of specific checkpoints that are bypassed in transformed cells and we are analysing these bypass mechanisms by comparative analysis of cell cycle regulation in normal fibroblasts versus transformed human cell lines.
Insulin Signaling and Akt kinase family
The Akt (protein kinase B) family of protein kinases are an integral part of insulin and other signalling cascades and have been implicated in essential cellular pathways and processes including cell differentiation and transformation. Our studies are focusing on differentiating potential interacting partners, such as p21cip and p57kip, and the specific action of Akt1 and Akt2 isoforms in proliferating normal or transformed cells. Our objectives are to identify key events and substrates for Akt involved in tumorigenesis and the epithelial-mesenchymal transition. In addition to bringing a better understanding of how Akt instigates tumorigenic changes, this will allow to develop isoform-specific tools with potential value as diagnostic markers and targets for therapy.

Muscle Derived Stem Cells, MDSC

Our second major research theme involves the isolation and characterization of a non-adherent population of skeletal muscle-derived stem cells, MDSC, capable of multipotent differentiation particularly into spontaneously beating cardiac muscle cells and neuronal lineages.
- Cell Therapy Potential of MDSC
In collaboration with IGF teams, we are analysing the in vivo multi-lineage differentiation and physiological repair potential of MDSC using mouse models of genetically- or chemically-targeted diseases and lineage-specific tracking of MDSC differentiation in particular towards cardiac and beta-pancreatic differentiation.
- Cardiac Rhythm disorders
Beating myocytes differentiated from MDSC in vitro are shown to be fully functional pacemaker myocytes such as those in the sino-atrial node (SAN) of the heart and transplantation experiments in mutant mice revealed that multipotent MDSC improved heart rhythm while engrafting into the SAN of severely bradycardic mice thus proving a very promising repair and regeneration potential.
- Insulin deficiency and diabetes.
In our on going work on MDSC, we are also investigating another differentiation lineage arising ex vivo from cultured MDSC and showing expression of several markers consistent with differentiation into beta pancreatic precursor cells. Our study shows that MDSC can differentiate into beta islet-like cell clusters in vitro and engraft to pancreatic islets in vivo where they differentiate into insulin-expressing cells in diabetic stretptozotocin-treated mice.
- Teratogenic potential and cancer stem cells.
Importantly, MDSC can be directly transplanted without inducing them into any differentiation lineage, being non tumorigenic up to several months after subcutaneous xenografting or intra peritoneal injection into immunodeficient mice. Without the need for induction into a pre-differentiated stage, MDSC direct use preserves the high plasticity, survival and migratory potential of transplanted cells. This is in sharp contrast with iPSC or ESC which develop fast growing lethal tumours 2-3 weeks after grafting if used without pre-differentiation. MDSC thus represent a valuable multivalent low risk source of autologous stem cells for regenerative therapy approaches.

Ned Lamb


Publications of the team

Regulation of the conserved 3'-5' exoribonuclease EXOSC10/Rrp6 during cell division, development and cancer.

Stuparević I, Novačić A, Rahmouni AR, Fernandez A, Lamb N, Primig M


Interactions between gut microbiota and skeletal muscle.

Gizard F, Fernandez A, De Vadder F

Metformin partially reverses the inhibitory effect of co-culture with ER-/PR-/HER2+ breast cancer cells on biomarkers of monocyte antitumor activity

Dahmani Z., Addou-Klouche L, Gizard F., Dahou S, Messaoud A, Chahinez Djebri N, Benaissti M.I, Mostefaoui M, Terbeche H, Nouari W, Miliani M, Lefranc . , Fernandez A, Lamb N J. and Aribi M.

Combination of metformin with sodium selenite induces a functional phenotypic switch of human GM-CSF monocyte-derived macrophages.

Meziane W, Mekkaoui Z, Hai I, Kacimi K, Djilali K, Touil-Boukoffa C, Lefranc G, Fernandez A, Lamb N, Mennechet F, Aribi M

Bradycardic mice undergo effective heart rate improvement after specific homing to the sino-atrial node and differentiation of adult muscle derived stem cells

Mesirca P., Mamaeva, D., Bidaud, I., Davase, R., DiFrancesco M.L., Mitutsova, V., Torrente, A.G., Arsic, N., Nargeot, J., Striessnig, J., Lee, A., Lamb, N., Mangoni, M., Fernandez A.

Adult Muscle-derived stem cells engraft and differentiate into insulin-expressing cells in pancreatic islets of diabetic mice

Mitutsova V, Wai-Yeng-Yeo W, Davaze R, Franckhauser C, Hani E, Abdullah S, Mollard P, Schaeffer M, Fernandez A and Lamb NJ.

Type II PKAs are anchored to mature insulin secretory granules in INS-1 β-cells and required for cAMP-dependent potentiation of exocytosis

Villalpando S, Cazevieille C, Fernandez A, Lamb NJ, Hani EH.

Cardiac arrhythmia induced by genetic silencing of 'funny' (f) channels is rescued by GIRK4 inactivation

Mesirca P, Alig J, Torrente AG, Müller JC, Marger L, Rollin A, Marquilly C, Vincent A, Dubel S, Bidaud I, Fernandez A, Seniuk A, Engeland B, Singh J, Miquerol L, Ehmke H, Eschenhagen T, Nargeot J, Wickman K, Isbrandt D, Mangoni ME.

The addressing fragment of mitogaligin: first insights into functional and structural properties

Senille V1, Lelievre D, Paquet F, Garnier N, Lamb N, Legrand A, Delmas AF, Landon C.

Characterization of the Akt2 Domain Essential for Binding Nuclear p21cip1 to Promote Cell Cycle Arrest during Myogenic Differentiation

Heron-Milhavet L, Franckhauser C, Fernandez A, Lamb NJ.

Purification and biochemical analysis of catalytically active human cdc25C dual specificity phosphatase

Franckhauser C, Fernandez A, Lamb NJ.

Akt1 and Akt2: Differentiating the aktion

Heron-Milhavet L, Khouya N, Fernandez A, Lamb NJ.

Distinct Pools of cdc25C Are Phosphorylated on Specific TP Sites and Differentially Localized in Human Mitotic Cells.

Franckhauser, C., Mamaeva, D., Heron-Milhavet, L., Fernandez, A., Lamb, NJ

Impaired muscle regeneration and myoblast differentiation in mice with a muscle-specific KO of IGF-IR

Heron-Milhavet L, Mamaeva D, Leroith D, Lamb NJ, Fernandez A.

Amazonia!: An Online Resource to Google and Visualize Public Human whole Genome Expression Data

Le Carrour T, Assou S, Tondeur S, Lhermitte L, Lamb Ned , Rème T, Pantesco V, Hamamah S, Klein B, De Vos J

Muscle-Derived Stem Cells isolated as Non-Adherent Population give rise to Cardiac, Skeletal Muscle and Neural Lineages.

Arsic, N., Mamaeva, D., Lamb, N.J., Fernandez, A.

The cell cycle control protein cdc25C is present and phosphorylated on serine 214 in the transition from germinal vesicle to metaphase II in human oocyte meiosis.

Cunat, S., Anahory, T., Berthenet, C., Hedon, B., Fernandez, A., Hamamah, S. and Lamb, NJ.

Akt2 is implicated in skeletal muscle differentiation and specifically binds Prohibitin2/REA

Heron-Milhavet L., Mamaeva D., Rochat A., Lamb NJC, and Fernandez A

Identification of a New Hybrid SRF and MEF2-binding Element in MyoD Enhancer Required for MyoD Expression during Myogenesis.

L'honore, A., Rana, V., Arsic, N., Franckhauser, C., Lamb, NJ., Fernandez, A.

Linking PCNA-dependent replication and ATR by human Claspin.

Brondello JM, Ducommun B, Fernandez A, Lamb NJ.

Only Akt1 is required for proliferation, while Akt2 promotes cell cycle exit through p21 binding

Héron-Milhavet L, Franckhauser C, Rana V, Berthenet C, Fisher D, Hemmings BA, Fernandez A, Lamb NJ.

Increased SP1 overall chromatin recruitment in divising cells by coordinated PP2A-mediated dephosphorylation

Vicart, A., Lefebvre, T., Imbert, J., Fernandez, A., Kahn-Perlès, B.

Only Akt1 is required for proliferation while Akt2 promotes cell cycle exit through p21 binding.

Heron-Milahvet, L., Franckhauser, C., Rana, V., Berthenet, C., Fisher, D., Hemmings, BA., Fernandez, A., Lamb, NJC.,

Comparative protein expression profiling in human cumulus cells in relation to oocyte fertilization and ovarian stimulation protocol

Hamamah, S., Matha, V., Berthenet, C., Anahory, T., Loup, V., Dechaud, H., Hedon, B., Fernandez, A., Lamb, NJC.

Subcellular localization and structural function of endogenous phosphorylated phosphatidylinosital 4-kinase (p14k92)

Szivak I, Lamb NJC, Heilmeyer LM.

Intracellular retention of Caveolin 1 in presenilin-deficient cells.

Wood D.R., Nye J.S., Lamb N.J., Fernandez A., Kitzmann M.

Increased levels of ANT1 protein and response to oxidative stress are early events in FSHD muscle

Laoudj-Chenivesse D., Carnac G., Bisbal C., Hugon G, Bouillot S, Desnuelle C, Vassetzky Y. and Fernandez A.

Insulin and Wnt1 pathways cooperate to induce reserve cell activation in differentiation and myotube hypertrophy

Rochat A., Fernandez A., Vandromme M, Moles JP, Bouschet T, Carnac G and Lamb N.J.C.

An Enhancer Directs Differential Expression of the Linked Mrf4 and Myf5 Myogenic Regulatory Genes in the Mouse

Chang T. H-T., Primig M., Hadchouel J., Tajbakhsh S., Rocancourt D., Fernandez A., Kappler R., Scherthan H. and M. Buckingham

MyoD Distal Regulatory Region contains an SRF binding CArG element required for MyoD expression in skeletal myoblasts and during muscle regeneration

L'honore A., Lamb N.J., Vandromme M., Turowski P., Carnac G. and Fernandez A

GermOnline, a new cross-species community annotation database on germ-line development and gametogenesis

M Primig, C Wiederkehr, R Basavaraj, C Sarrauste de Menthière, L Hermida, R Koch, U Schlecht , HG Dickinson, M Fellous, JAGrootegoed 5 , R S Hawley, B Jégou, B Maro, A Nicolas, T Orr-Weaver, T Schedl, A Villeneuve, DJ Wolgemuth, M Yamamoto D Zickler

Transient induction of cyclin A in loaded chicken skeletal muscle.

Fluck M, Kitzmann M, Dapp C, Chiquet M, Booth FW and Fernandez A

Functional cdc25C dual-specificity phosphatase is required for S-phase entry in human cells

Turowski P., Franckhauser C., Morris, CM, Vaglio P., Fernandez A. and Lamb N.J.C.

HIV-1 Tat-associated CTD kinase, CDK2, phosphorylates CDK7 and stimulates Tat-mediated transcription.

Nekhai S., Zhou M., Fernandez A., Lane WS, Lamb N.J.C., Brady, J. and Kumar A.

Identification of new proteins in follicular fluid of mature human follicles

Tal Anahory, Hervé Dechaud, René Bennes, Philippe Marin, Ned J. Lamb and Dalila Laoudj

Macromolecular uptake is a spontaneous event during mitosis in cultured fibroblasts: implications for vector-dependent plasmid transfection

Pellegrin P, Fernandez A, Lamb N.J., Bennes R.

Crosstalk between cell cycle regulators and the myogenic factor MyoD in skeletal myoblasts.

Kitzmann, M., Fernandez, A.

Protein kinase B beta/Akt2 plays a specific role in muscle differentiation

Marie Vandromme, Anne Rochat, Roger Meier, Gilles Carnac, Daniel Besser, Brian A. Hemmings, Anne Fernandez, and Ned J. C. Lamb

Cell cycle-dependent stimulation of the HIV-1 promoter by tat-associated CAKactivator.

Nekhai S, Shukla RR, Fernandez A, Kumar A, Lamb NJ

Mutation of MyoD-Ser237 abolishes its up-regulation by c-Mos

Pelpel K, Leibovitch M, Fernandez A, Leibovitch SA.

Cyclin E-Cdk2 Phosphorylation Promotes Late G1-Phase Degradation of MyoD in Muscle Cells.

Tintignac LA, Leibovitch MP, Kitzmann M, Fernandez A, Ducommun B, Meijer L, Leibovitch SA.

The retinoblastome-like p130 is involved in the determination of reserve cells in differentiating myoblasts.

Carnac, G., Fajas, L., L'Honoré, A., Sardet, C., Lamb, N.J.C., and Fernandez, A.

CHF: a novel factor binding to cyclin A CHR corepressor element.

Philips A, Chambeyron S, Lamb N, Vie A, Blanchard JM

Vimentin dephosphorylation by protein phosphatase 2A is modulated by the targetting subunit B55.

Turowski, P., Myles, T., Hemmings, B.A., Fernandez, A. and Lamb, N.

Cdk1- and cdk2-mediated phosphorylation of MyoD ser200 in growing C2 myoblasts: role in modulating MyoD half-life and myogenic activity.

Kitzmann, M., Vandromme, M., Shaeffer, V., Carnac, G., Labbé, J.C., Lamb, N. and Fernandez, A.

The structure of the protein phosphatase 2A PR65/A subunitreveals the conformation of its 15 tandemly repeated HEAT motifs.

Groves, R.M., Hanion, N., Turowski, P., Hemmings, B.A. and Barford, D.

Inhibition of mammalian cell proliferation by genetically selected peptide aptamers that functionnaly antagonize E2F activity.

Fabbrizio, E., Le Cam, L., Polanowska, J., Kaczorek, M., Lamb, N., Brent, R. and Sardet, C.

Functional specificity of the two retinoic acid receptor RAR and RXR families in myogenesis.

Alric, S., Froeschlé, A., Piquemal, D., Carnac, G, and Bonnieu, A.

Un peptide synthétique vecteur pour les rétinoïdes et anti-prolifératif.

Pellegrin, P., Mery, J. and Bennes, R.

Microinjection and immunological methods in the analysis of type 1 and 2A protein phosphatase from mammalian cells.

Turowski, P. and Lamb, N.

Aggregates of an amphiphilic synthetic peptide bind and deliver all-trans retinol and all-trans retinoic acid into fibroblasts cells.

Pellegrin, P., Chaloin, L., Roustan, C., Méry, J. and Bennes, R.

The muscle regulatory factors MyoD and Myf-5 undergo distinct cell-specific expression in muscle cells.

Kitzmann, M., Carnac, G., Vandromme, M., Primig, M., Lamb, N. and Fernandez, A.

Retinoïc acid receptor and muscle bHLH partnersin retinoid induced myogenesis.

Froeschlé, A., Alric, S., Kitzmann, M., Carnac, G., Auradé, F., Rochette-Egly, C. and Bonnieu, A.

Molecular cloning, chromosomal localisation and cell cycle-dependent subcellular distribution of the A-kinase anchoring protein, AKAP95.

Eide, T., Coghlan, V., Orstavik, S., Holsve, C., Solberg, R., Skalhegg, B., Lamb, N., Langeberg, L., Fernandez, A., Scott, J., Jahnsen, T. and Tasken, K.

RhoA GTPase and serum response factor control selectively the expression of MyoD without affecting Myf5 in mouse myoblasts.

Carnac, G., Primig, M., Kitzmann, M., Chafey, P., Tuil, D., Lamb, N. and Fernandez, A.

Rôle of the multi functional Trio protein in the control of the Rac1 and RhoA GTPase signaling pathways.

Bellanger, J.M., Zugasti, O., Lazaro, J.B., Diriong, S., Lamb, N., Sardet, C. and Debant, A.

The two guanine nucleotide exchange factor domains of Trio link the Rac1 and RhoA pathways in vivo.

Bellanger, J.M., Lazaro, J.B., Diriong, S., Fernandez, A., Lamb, N. and Debant, A.

Thèses et hdr

The role of the HSP90/R2TP system in the assembly of the TSC complex and the regulation of the mTORC1 pathway 16/07/2021

Defended by Claire Abeza on 10/06/2021