In-Vitro 2D & 3D Neuronal Networks Summit

Speaker

Prof. Nael Nadif Kasri
Radboud University Medical Centre, Department of Human Genetics,Donders Institute for Brain, Cognition and Behaviour, Netherlands

Bio

Nael Nadif Kasri did his PhD in molecular biology at the KU Leuven. After his PhD, he worked as a postdoctoral researcher in neurobiology at CSHL, New York. In 2010 he started his independent research group at the Radboudumc, where he is part of the Donders institute. The focus of his research is to understand the synaptic basis of neurodevelopmental disorders. In particular, his expertise in the generation of patient-derived neural lineages and their neurophysiological analysis at single cell and network level (MEA) is setting the stage in this novel research field.

Abstract

Recent progress in human genetics has led to the identification of hundreds of genes associated with neurodevelopmental disorders. In his talk, Nael Nadif Kasri will discuss his strategy to link genetic deficits observed in patients to neuronal network measurements. He combines induced pluripotent stem cell-derived neurons (excitatory and inhibitory) with micro-electrode arrays (MEAs) and transcriptomics analysis (MEA-seq) to unravel the pathomechanism underlying specific syndromes and identify genotype-phenotype correlations.

Speaker

Prof. Andreas Hierlemann
Bio Engineering Laboratory, D-BSSE, ETH Zurich, Basel, Switzerland

Bio

Andreas Hierlemann got his college education in chemistry at the University of Tübingen, Germany and a Ph.D. degree in 1996. He held Postdoc positions 1997 at Texas A & M University, College Station, TX and 1998 at Sandia National Laboratories, Albuquerque, NM, USA. He joined the Department of Physics of ETH Zurich 1999, where he was appointed Associate Professor in 2004. In 2008, he became Full Professor in the Department of Biosystems Science and Engineering of ETH Zurich in Basel.

Abstract

High-density MEAs feature thousands of densely packed small electrodes (>3000 electrodes per mm2 with diameters of 5-7 µm and pitch of <15 µm) and can be used for interaction with networks of, e.g., neurons at cellular or subcellular resolution in dissociated cell cultures, organotypic and acute tissue slices. Complementary metal-oxide-semiconductor (CMOS)-technology is used to batch-produce those complex microsystems, which feature – on the very same chip – dense arrays of tens of thousands of electrodes along with addressing logic and circuitry units for signal conditioning.

Topic | Functional Characterization of Human iPSC-derived Neurons – Technologies and Challenges

Panelists:
1. Dr. Kenta Shimba, Group of Prof. Yasuhiko Jimbo, University of Tokyo, Japan

Bio | Kenta Shimba has been working as an assistant professor at Professor Jimbo’s laboratory in the University Tokyo since 2018. His areas of expertise include microdevice fabrication which can combine with microelectrode array, and evaluation of axon conduction characteristics and iPS cell-derived neurons.

2. Prof. Nael Nadif Kasri, Radboud University Medical Centre, Department of Human Genetics, Donders Institute for Brain, Cognition and Behaviour, Netherlands

Bio | Nael Nadif Kasri did his PhD in molecular biology at the KU Leuven. After his PhD, he worked as a postdoctoral researcher in neurobiology at CSHL, New York. In 2010 he started his independent research group at the Radboudumc, where he is part of the Donders institute. The focus of his research is to understand the synaptic basis of neurodevelopmental disorders. In particular, his expertise in the generation of patient-derived neural lineages and their neurophysiological analysis at single cell and network level (MEA) is setting the stage in this novel research field.

3. Dr. Bruno Buisson, Neuroservices-Alliance & Neuroservice, USA & France

Bio | Bruno has a long/strong background in neurosciences/electrophysiology. Collaborating with Pharma/biotech companies for more than 25 years, he has extensively used electrophysiological techniques and assays to investigate compounds’ pharmacological properties on neurons and neuronal networks. As a post-doc he has “played” with the first MEA systems commercially available in the mid of the 90’s by MSC. With Olivier TOURY he launched NEUROSERVICE in 2006.

4. Dr. Silvia Ronchi, Group of Prof. Andreas Hierlemann, Bio Engineering Laboratory, D-BSSE, ETH Zurich, Basel, Switzerland

Bio | Silvia Ronchi received the B.Sc. degree in clinical engineering in 2014 and the M.Sc. degree in biomedical engineering in 2017 from Università degli Studi di Roma “La Sapienza”, Italy. Since 2017, she has been working at the Bio Engineering Laboratory, ETH Zürich, Basel, Switzerland, where she recently finished her PhD studies. Her current research interests include the electrophysiological phenotype characterization of human iPSC-derived neurons modeling neurodegenerative diseases.

5. Dr. Carsten Pfeffer, Neurolentech GmbH, IST Austria Science Park, Austria

Bio | Carsten received his PhD Suma Cum Laude from the Centre for Molecular Neurobiology, from Berlin, Germany, in 2008. He held Postdoc positions in 2008 in the lab of Thomas Jentsch at MDC and FMP in Berlin and from 2010 until 2017 in the lab of Massimo Scanziani, HHMI/UCSD in California. Carsten was awarded with an EMBO Fellowship from 2010 until 2012 and an NIH BRAIN award on Census of cell-types in 2014. Carsten is the Co-founder and CEO of Neurolentech GmbH, an IST Austria Spin-off company: personalized medicine for Autism and Epilepsy.

Speaker

Martina de Gennaro
Group of Prof. Botond Roska, Institute of Molecular and Clinical Ophthalmology Basel, University of Basel, Switzerland

Bio

Martina attained her B.Sc. degree in Biotechnology “cum Laude” from University La Sapienza in Rome, with an experimental thesis in the field of Immunology and Immunopathology. She further pursued her studies at ETH Zurich, carrying out research projects in both molecular and clinical neurobiology. She attained her M.Sc. degree in Biology, majoring in Neuroscience, defending a thesis on the development of inhibitory synaptic circuits. Martina is currently a doctoral candidate at the Institute of Ophthalmology Basel, mentored by Professor Roska. Her research focuses on the functional characterization of retinal ganglion cells across species.

Abstract

Light-sensitive human retinal organoids recapitulate cell-types, multilayering and transcriptomic profiles of the human retina, offering a relevant tool for translational studies. The generation of a library of 285,441 single-cell transcriptomes from light-responsive human retinae and retinal organoids at different time points allows the comparison of developmental rates and genomic expression profiles. Retinal organoids converge to adult peripheral retinal cell-type-specificity at a pace comparable to human retina development in vivo. Electrophysiological assessments, such as multi-electrode-array and calcium imaging, ensure the collection of transcriptomes from functional, light-responsive human retinae ex-vivo and serve to identify synaptic-driven light-sensitivity in mature human retinal organoids. Moreover, expression of disease-associated genes in the human retina is preserved in mature retinal organoids, allowing to define cellular targets for disease mechanisms investigation in organoids and targeted repair in the human retina.

Speaker

Ana Rita Gomes
Group of Prof. Joaquim M. Sampaio Cabral, Dept. of Bioengineering, IBB and i4HB, Instituto Superior Técnico, University of Lisbon, Portugal

Bio

Ana Rita Gomes is a PhD candidate in Bioengineering – Cell Therapies and Regenerative Medicine at Instituto Superior Técnico, University of Lisbon. Ana Rita has her MSc in Biological Engineering, Instituto Superior Técnico, University of Lisbon. Her research is focused on the study of organoid biology and process engineering strategies that combined to develop region specific brain organoids, mainly containing ventral and dorsal forebrain characteristic neurons, from human induced pluripotent stem cells (hiPSCs). The main objective is to use the organoids platform for modeling a neurodevelopmental disorder, Rett Syndrome (RTT).

Abstract

Engineering brain organoids from human induced pluripotent stem cells (hiPSCs) is a powerful tool for modeling brain development and neurological disorders. Rett Syndrome (RTT), a rare neurodevelopmental disorder, can greatly benefit from this technology, since it affects multiple neuronal subtypes in forebrain sub-regions. We have established dorsal and ventral forebrain organoids from control and RTT patient-specific hiPSCs recapitulating 3D organization and functional network complexity. Our data revealed a premature development of the deep-cortical layer, associated to the formation of TBR1 and CTIP2 neurons, and a lower expression of neural progenitor/proliferative cells in female RTT (MeCP2:R255X) dorsal organoids. Moreover, calcium imaging and electrophysiology analysis demonstrated functional defects of RTT neurons. Additionally, assembly of RTT dorsal and ventral organoids revealed impairments of interneuron’s migration. On the other side, RTT male (MeCP2:Q83X) interneurons derived from ventral organoids exhibited consistently defaults in spike activity and in synaptic maturation.
Overall, our models provide a better understanding of RTT during early stages of neural development, demonstrating a great potential for personalized diagnosis and drug screening.

Speaker

Dr. Silvia Benito-Kwiecinski
Group of Prof. Madeline Lancaster, MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, England

Bio

Silvia is currently a postdoc in Dr Madeline Lancaster’s laboratory at the MRC Laboratory of Molecular Biology where she recently completed her PhD with the University of Cambridge. Silvia is interested in neurodevelopment and evolution. Her research involves comparing early development of brain organoids derived from human and non-human ape species to identify and study human-specific features of brain development.

Abstract

Human evolution has been marked by a rapid expansion in brain size since our divergence from other apes. Brain organoids model the earliest stages of brain development and, by generating human, gorilla and chimpanzee derived organoids, we can study how and when size differences begin to arise. We have found that human organoids become larger than the other apes prior to the formation of neurons and result from a human delay in the transition of neural progenitor cells from proliferative to neurogenic. We have identified ZEB2 as a regulator of this progenitor cell transition.

Speaker

Prof. Kenneth Kosik
Neurobiology Lab, UC Santa Barbara, USA

Bio

Kenneth S. Kosik completed a B.A. and M.A. in English literature from Case Western Reserve University in 1972 and an M.D. from the Medical College of Pennsylvania in 1976. He served as a resident in neurology at Tufts New England Medical Center and was Chief Resident there in 1980. Beginning in 1980 he held a series of academic appointments at the Harvard Medical School and achieved the rank of full professor there in 1996. He also held appointments at McLean Hospital, Brigham and Women’s Hospital, the Massachusetts General Hospital and the Dana-Farber Cancer Institute. In 2004, Kosik became the Harriman Professor of Neuroscience Research and Co-Director of the Neuroscience Research Institute at the University of California, Santa Barbara. He is the recipient of the 2021 Potamkin Prize for Alzheimer’s disease research and is a fellow of the American Association for the Advancement of Science.

Abstract

Human brain organoids replicate much of the cellular diversity and developmental anatomy of the human brain. However, the physiological behavior of neuronal circuits within organoids remains relatively under-explored. With high-density CMOS microelectrode arrays and shank electrodes, we probed broadband and three-dimensional spontaneous activity of human brain organoids. These recordings simultaneously captured local field potentials (LFPs) and single unit activity. From spiking activity, we estimated a directed functional connectivity graph of synchronous neural network activity which showed a large number of weak functional connections enmeshed within a network skeleton of significantly fewer strong connections. Increasing the intrinsic inhibitory tone with a benzodiazepine altered the functional network graph of the organoid by suppressing the network skeleton. Simultaneously examining the spontaneous LFPs and their phase alignment to spiking showed that spike bursts were coherent with theta oscillations in the LFPs. An ensemble of spikes phase-locked to theta frequency oscillations were strongly interconnected as a sub-network within the larger network in which they were embedded. Our results demonstrate that human brain organoids have self-organized neuronal assemblies of sufficient size, cellular orientation, and functional connectivity to co- activate and generate field potentials from their collective transmembrane currents that phase- lock to spiking activity. These results point to the potential of brain organoids for the study of neuropsychiatric diseases, drug mechanisms, and the effects of external stimuli upon neuronal networks.

Topic: Current advancements in brain organoid / spheroid research and applications

Panelists:
1. Martina de Gennaro, Group of Prof. Botond Roska, Institute of Molecular and Clinical Ophthalmology Basel (IOB), Switzerland

Bio | Martina attained her B.Sc. degree in Biotechnology “cum Laude” from University La Sapienza in Rome, with an experimental thesis in the field of Immunology and Immunopathology. She further pursued her studies at ETH Zurich, carrying out research projects in both molecular and clinical neurobiology. She attained her M.Sc. degree in Biology, majoring in Neuroscience, defending a thesis on the development of inhibitory synaptic circuits. Martina is currently a doctoral candidate at the Institute of Ophthalmology Basel, mentored by Professor Roska. Her research focuses on the functional characterization of retinal ganglion cells across species.

2. Ana Rita Gomes, Group of Prof. Joaquim M. Sampaio Cabral, Department of Bioengineering and IBB-Institute for Bioengineering and Biosciences and Associate Laboratory i4HB-Institute for Health and Bioeconomy, Instituto Superior Técnico, University of Lisbon, Portugal

Bio | Ana Rita Gomes is a PhD candidate in Bioengineering – Cell Therapies and Regenerative Medicine at Instituto Superior Técnico, University of Lisbon. Ana Rita has her MSc in Biological Engineering, Instituto Superior Técnico, University of Lisbon. Her research is focused on the study of organoid biology and process engineering strategies that combined to develop region specific brain organoids, mainly containing ventral and dorsal forebrain characteristic neurons, from human induced pluripotent stem cells (hiPSCs). The main objective is to use the organoids platform for modeling a neurodevelopmental disorder, Rett Syndrome (RTT).

3. Dr. Silvia Benito-Kwiecinski, Group of Dr. Madeline Lancaster, MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, England

Bio | Silvia is currently a postdoc in Dr Madeline Lancaster’s laboratory at the MRC Laboratory of Molecular Biology where she recently completed her PhD with the University of Cambridge. Silvia is interested in neurodevelopment and evolution. Her research involves comparing early development of brain organoids derived from human and non-human ape species to identify and study human-specific features of brain development.

4. Prof. Kenneth Kosik, Neurobiology Lab, UC Santa Barbara, USA

Bio | Kenneth S. Kosik completed a B.A. and M.A. in English literature from Case Western Reserve University in 1972 and an M.D. from the Medical College of Pennsylvania in 1976. He served as a resident in neurology at Tufts New England Medical Center and was Chief Resident there in 1980. Beginning in 1980 he held a series of academic appointments at the Harvard Medical School and achieved the rank of full professor there in 1996. He also held appointments at McLean Hospital, Brigham and Women’s Hospital, the Massachusetts General Hospital and the Dana-Farber Cancer Institute. In 2004, Kosik became the Harriman Professor of Neuroscience Research and Co-Director of the Neuroscience Research Institute at the University of California, Santa Barbara. He is the recipient of the 2021 Potamkin Prize for Alzheimer’s disease research and is a fellow of the American Association for the Advancement of Science.

5. Dr. Tal Sharf, Group of Prof. Kenneth Kosik, Neurobiology Lab, UC Santa Barbara, USA

Bio | Utilizing techniques at the intersection of physical science and biology, he is developing new techniques to investigate neural circuitry utilizing human brain organoids with the aim to uncover general rules to explain how they malfunction with disease and mental illness.