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Understanding single cell-level MAPK activation dynamics for manipulation of neuronal stem cell self-renewal and differentiation fates

English title Understanding single cell-level MAPK activation dynamics for manipulation of neuronal stem cell self-renewal and differentiation fates
Applicant Pertz Olivier
Number 162195
Funding scheme Bilateral programmes
Research institution Departement Biomedizin Universität Basel
Institution of higher education University of Berne - BE
Main discipline Neurophysiology and Brain Research
Start/End 01.03.2016 - 28.02.2019
Approved amount 250'000.00
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Keywords (5)

neuronal differentiation; MAP Kinase; neural stem cell; microfluidics; live cell imaging

Lay Summary (German)

Lead
Signalisierungsprozesse die Stammzellen Differenziation modulieren
Lay summary

Neue Fortschritte in der Stammzellenforschung ermöglichen die Herstellung von einer großen Anzahl von spezifischen Zelltypen für Zellersatztherapie oder Krankheitsforschung. Limitierende Faktoren sind die tiefe Effizienz mit der der gewünschte Zelltyp produziert werden kann, sowie die vielen verschiedenen Zelltypen die typischerweise in einer Zellpopulation entstehen. Die Fähigkeit einer undifferenzierten Stammzelle in einen spezifischen Zelltyp zu differenzieren ist von komplexen Signal Ereignisse geregelt. Es wurde kürzlich gezeigt, dass die Zellen innerhalb einer Population sehr unterschiedliche Signale aufweisen können. Dies erklärt die beobachtete Zellschicksal Heterogenität. Vor kurzem, haben wir mit einer Kombination von System Biologie und Engineering gezeigt wie man die Signaldynamik in einer Zellpopulation homogenisiert. Dadurch können sehr homogene Zellschicksale innerhalb einer Zellpopulation induziert werden. In diesem Projekt wollen wir mit ähnliche Ansätzen neurale Stammzellen behandeln um eine effektive Produktion von homogenen Nervenzellen zu produzieren.

 

Direct link to Lay Summary Last update: 04.03.2016

Responsible applicant and co-applicants

Employees

Name Institute

Publications

Publication
Temporal perturbation of ERK dynamics reveals network architecture of FGF2/MAPK signaling.
(2019), Temporal perturbation of ERK dynamics reveals network architecture of FGF2/MAPK signaling., in Molecular Systems Biology, 15(11), e8947.
Integrated Platform for Monitoring Single-cell MAPK Kinetics in Computer-controlled Temporal Stimulations
Ryu Hyunryul, Chung Minhwan, Song Jiyoung, Lee Sung Sik, Pertz Olivier, Jeon Noo Li (2018), Integrated Platform for Monitoring Single-cell MAPK Kinetics in Computer-controlled Temporal Stimulations, in Scientific Reports, 8(1), 11126-11126.
Microfluidic platform for single cell analysis under dynamic spatial and temporal stimulation
Song Jiyoung, Ryu Hyunryul, Chung Minhwan, Kim Youngtaek, Blum Yannick, Lee Sung Sik, Pertz Olivier, Jeon Noo Li (2018), Microfluidic platform for single cell analysis under dynamic spatial and temporal stimulation, in Biosensors and Bioelectronics, 104, 58-64.

Collaboration

Group / person Country
Types of collaboration
Prof. Noo Li Jeon, Seoul National University Korean Republic (South Korea) (Asia)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Exchange of personnel
Verdon Taylor, University of Basel Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
Goodbye Flat Biology: In Vivo inspired Cancer Biology and Therapy Poster ERK and Akt Signalling Dynamics in 3D Organoid Models of Breast Cancer - Associated Mutations 09.09.2018 Berlin, Germany Ender Pascal;
BeFri Research Retreat 2018 Poster ERK and Akt Signalling Dynamics in 3D Organoid Models of Breast Cancer - Associated Mutations 26.04.2018 kandersteg, Switzerland Ender Pascal;
GCB Symposium 2018 Poster Understanding Single Cell - Level MAPK Activation Dynamics for Manipulation of Neuronal Stem Cell Self - Renewal and Differentiation Fates 01.02.2018 bern, Switzerland Ender Pascal;
LS2 meeting Poster Understanding single c ell - level MAPK activation dynamics for manipulation of neuronal stem cell self - renewal and differentiation fates 02.02.2017 Zurich, Switzerland Jeon Noo Li; Pertz Olivier; Ender Pascal;


Associated projects

Number Title Start Funding scheme
185376 Decoding and Re-Encoding Receptor Tyrosine Kinase/Fate Decision Signaling 01.06.2019 Project funding (Div. I-III)
149923 Optogenetic control of receptor tyrosine kinase signaling to manipulate cell fate 01.02.2014 Project funding (Div. I-III)
176008 Hunting for natural products targeting aberrant proliferative signaling in melanoma 01.01.2018 Project funding (Div. I-III)

Abstract

Recent advances in stem cell research enable generation of large number of specific cell types for cell-replacement therapy or disease research. One available approach consists of “in-vitro directed differentiation” in which addition of growth factors (GFs) or small molecules are used to mimic specific steps occurring during embryonic development. These GFs activate signaling events that control the gene expression programs inducing self-renewal or differentiation to different cell types. Empirical approaches have led to a plethora of protocols in which GF/small molecule combinations enable to control self-renewal capability or differentiation. An important current limitation is that the efficiency with which a desired cell fate is specified, remains low, and many different cell lineages are specified within the cell population. This is thought to arise from heterogeneous signaling states in the population. Understanding this signaling heterogeneity, and how it relates to self-renewal or differentiation fates at the single cell level might provide novel leads for robust control of self-renewal capability and differentiation to specific fates.Recent technological advances in single cell analysis enable investigation of heterogeneous signaling states within a cell population that are not accessible using classic population average measurements such as Western Blots. Advanced imaging and analysis of single cell behavior provides a conceptual framework to understand heterogeneous cell fate specification. Furthermore, signaling dynamics rather than steady states have recently been shown to be important for cell fate determination. ERK MAPKs are key regulators of cell fate decisions such as differentiation, proliferation and death, and have been involved in stem cell biology. The Pertz and Jeon lab have recently studied the effect of temporal GF stimulation on single cell ERK MAPK dynamics to control cell fate in cultured PC-12 cell model system. Two distinct GFs (EGF and NGF) can control the duration of ERK MAP kinase activity to specify proliferation or neuronal differentiation. Taking advantage of a combination of FRET-based sensors to measure ERK activation dynamics in single living cells, and microfluidic technology to dynamically probe the MAPK network using pulsed GF stimulation (manuscript under review), our data explains how distinct signaling states arise in a cell population, and correlate with heterogeneous cell fates. Mathematical modeling, and its experimental verification, then showed that dynamic control of ERK MAPK dynamics, by temporally controlled delivery of GFs, can induce robust and homogenized signaling states throughout the population. As a consequence, dynamic control of ERK activity leads to more robust and homogeneous cell fate induction within the population, independently of GF identity.In this proposal, we propose to extend our novel conceptual framework to a primary mouse stem cell system: dentate gyrus adult neural stem cells (DG-aNSCs). We have access to reagents, technologies and knowledge about this model system through collaboration with the lab of Verdon Taylor in our current department at the University of Basel. The Pertz lab has already started to use these cells in the context of another neurobiology-related project. Understanding single cell MAPK dynamics in the control of self-renewal and differentiation in these DG-aNSCs might provide novel ways to induce robust differentiation of these stem cells into hippocampal neurons. Specifically, we will build a DG-aNSC cell line in which proliferation (cell cycle entry) or differentiation fate (into the neuronal lineage), as well as ERK activation dynamics can be dynamically measured in a multiplexed fashion in the same cell. We will use state of the art long term time lapse imaging (on a timescale of multiple days), to relate ERK activation dynamics to self-renewal proliferative or neuronal differentiation cell fates. This will be performed under experimental conditions that induce stem cell renewal, or neuronal differentiation, which both require specific GFs that are known to activate the ERK MAPK pathway. Once we have related dynamic ERK signaling patterns to self-renewal or differentiation fates, we will design dynamic GF stimulation schemes that closely mimic these specific dynamics with the aim to induce robust fates. As we have already shown in PC-12 cells, this might have the potential to homogenize the signaling state, and thus the cell fate within the cell population. An important current limitation is that our microfluidic approaches necessary to dynamically deliver GF pulses for now only enables manipulation of a small cell population for analytical purposes. We therefore propose to evolve this technology to scale the cell number up for preparative purposes, that could then be employed for disease research. Our multidisciplinary approach will provide novel insights into the basic biology of the signaling dynamics that control stem cell self-renewal and differentiation at the single cell level in a biologically and disease-relevant model system, and might yield novel ways to robustly manipulate both processes.
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