Center for Human-Intelligence Research

Director: Hideyuki Okano (Professor, School of Medicine)

Campus: Shinanomachi

Cognitive science, brain science, robotics, society, civilization

■ Activities Continuing from FY2019: Background, Rationale, and Goals

Following the previous fiscal year, the center's groups—"Elucidation from Molecular Biology and Developmental Engineering," "Elucidation from Comparative Cognitive Science," "Elucidation from Brain Science," and "Elucidation from Robotics"—will continue to work together in unison. They will explore human intelligence broadly, from the molecular to the behavioral level, through approaches not only from the natural sciences but also from the humanities and social sciences. The project items are as follows.

(01) In research on inducing human cognitive evolution through the interaction of environment, genes, and neural activity, we will continue to conduct behavioral and imaging analyses (MRI, PET) using genetically modified marmosets, which are primate models of disease, to detect behavioral abnormalities related to psychiatric and neurological disorders from an early stage, and prepare to publish the results.

For items (02) through (07) from the FY2019 Business Report, we will aim to publish them as papers or establish a clear path toward publication as significant achievements related to human intelligence by the final fiscal year.

(07) In the development of marmoset brain function mapping technology using fMRI, we will conduct detailed analyses of the relationship between marmoset gene expression and functionally connected regions.

(08) As part of the development of brain function mapping technology using electron microscopy, we will work on developing a method to visualize neural activity with the world's fastest multi-beam scanning electron microscope.

(09) For cell-type-specific long-term imaging of the marmoset brain using fluorescent calcium imaging technology, we will use an ultra-compact two-gram fluorescence microscope to develop technology for deep-brain function mapping in a free-moving environment.

(10) We will conduct correlative analysis using fluorescence and electron microscopy at the center. We will compile a paper on the analysis results regarding the characteristic axonal degeneration and the localization of molecules related to early lesions of Alzheimer's disease in Alzheimer's model mice, using cell and tissue labeling techniques that allow for comparative analysis with electron and light microscopy, and employing the world's fastest multi-beam scanning electron microscope.

(11) For the analysis of postmortem human brains with Alzheimer's disease, our policy is to obtain brains from Alzheimer's patients and conduct analyses of axonal degeneration and the localization of molecules related to the disease's early lesions.

(12) In the project to clarify the characteristics of human intelligence by comparing the neural mechanisms of spatial cognition in eels with those in humans, we will identify global cues for spatial cognition and the selection of these cues, as well as consider sensory deprivation and brain lesion exchanges in fish.

■ New Activity Goals and Content for FY2020, and Background for Implementation

We will begin the following new items from the next fiscal year.

(13) We will conduct research aimed at elucidating the complex motor control mechanisms in the striatum using deep-brain fluorescent calcium imaging technology.

(14) We will induce nervous system cells from patient-derived iPS cell lines or genome-edited lines and conduct research to reproduce epilepsy-like symptoms in vitro and study the pathogenic mechanisms of Alzheimer's disease using human cells.

■ Implementation Details, Research Results, and Degree of Achievement for the Fiscal Year's Business Plan

The research results for FY2019 are described for each research item.

(01) In research on inducing human cognitive evolution through the interaction of environment, genes, and neural activity, we are continuing to conduct behavioral analyses, such as sleep behavior, and imaging analyses (MRI, PET) using genetically modified model marmosets of psychiatric and neurological disorders to detect related behavioral abnormalities from an early stage.

(02) In the development of human disease model marmosets, we are conducting histological analysis of transgenic marmosets with forced expression of alpha-synuclein to continuously analyze early-onset changes in Parkinson's disease. Through analysis that captures disease-related changes early, we have been able to identify disease-specific impairments and will submit a paper.

(03) In the development of transgenic marmosets with human-specific genes, we are conducting phenotype analysis using the brains of mice with modified human-specific genes. In particular, we quantitatively analyzed the thickness of the cerebral cortex, which is specifically enlarged in the human brain. We have also succeeded in creating individual marmoset models with modified human-specific, developmentally important genes and are conducting histological analysis.

(04) To elucidate the neural basis of social behavior through communication, we are proceeding with the creation of autism model marmosets using new genome-editing technology.

(05) In the development of new methods for creating genetically modified primate animals, we have succeeded in increasing the efficiency of gene modification by applying new genome-editing technology to marmoset fertilized eggs, and we are currently in the process of creating genetically modified individuals.

(06) As part of comparative cognitive science research to identify the characteristics of human intelligence, we have developed a new method for quantitatively evaluating marmoset behavior over time using a new system for behavioral analysis of experimental animals, and this analysis is ongoing.

(07) In the development of brain function mapping technology using fMRI, we are applying the fMRI brain function mapping technology conducted in an awake state to perform fMRI analysis on genetically modified animals.

(08) As part of the development of marmoset brain function mapping technology using electron microscopy, we have succeeded in completing a function mapping method by enabling the acquisition of serial section images over a vast area with a multi-beam scanning electron microscope capable of high-speed imaging over an ultra-wide range.

(09) For cell-type-specific long-term imaging of the marmoset brain using fluorescent calcium imaging technology, we have succeeded in visualizing neural activity with fluorescent probes in free-moving marmosets using the recently developed two-gram ultra-compact fluorescence microscope, nVista, and are conducting task-based imaging.

(10) We conducted correlative analysis using fluorescence and electron microscopy at the center. This fiscal year, by applying tissue labeling techniques usable for both electron and light microscopy, we published a paper on some of our findings regarding the characteristic axonal degeneration and the localization of molecules related to early lesions of Alzheimer's disease in Alzheimer's model mice. We are generally on track with our plan, achieving our goals and obtaining results.

(11) For the analysis of postmortem human brains with Alzheimer's disease using electron microscopy to study axonal degeneration and the localization of molecules related to early lesions, we contacted the person in charge of the human brain bank, determined the brain regions, prepared the necessary documents, and completed the procedures.

(12) Regarding the experiment to clarify the characteristics of human intelligence by comparing the neural mechanisms of spatial cognition in eels with those in humans, we established an experimental system for eels and reported the results in the journal *Animal Cognition*.

Number of published papers: 18

01. Tanaka H et al. Nat Commun. 2020 in press.

02. Khazaei M et al. Sci Transl Med. 2020

03. Saeki T, Hosoya M, Shibata S et al. Neurosci Lett. 2020

04. Mohan S et al. J Neuropathol Exp Neurol. 2019

05. Serizawa T et al. Development. 2019

06. Ariyasu D et al. Endocrinology. 2019

07. Hoshino Y et al. Scientific Reports. 2019

08. Li Y, Kanzaki S, Shibata S et al. Neurosci Res. 2019

09. Shibata S, et al. Front Neural Circuits, 2019

10. Kirihara T, et al. iScience 2019

11. Yamazaki Y, Abe Y, Shibata S, et al. J Neurosci. 2019

12. Kinugasa Y, et al. Journal of Cell Science, 2019

13. Abe Y et al. Neurochem Int. 2019

14. Miki F, et al. Biol Reprod. 2019

15. Watanabe,S. Learning and Motivation, 2019

16. Watanabe S & Shinozuka K, Animal Cognition 2020

17. Watanabe S Learning and Behavior (in press)

18. Shigeru Watanabe, "Do Animals Need a Mind? Confronting Anthropomorphism" (University of Tokyo Press)

Number of invited lectures at academic conferences, etc. (domestic and international): (5 domestic, 1 international)

Achievements in social contribution, such as events: 3

01. "Brain Gakumon no susume (An Encouragement of Learning)," World Brain Week 2019, Keio University Shinanomachi Campus (a tour for high school students and others from across the country where researchers introduce the front lines of brain research, hosted by the Japan Brain Century Promotion Conference) 2019/11/2

02. "Let's Explore the Body with the World's Fastest Electron Microscope," Keio University Shinanomachi Campus (a laboratory tour during the school festival for local residents, high school students, etc.) 2019/10/19

03. Edogawa Library Civic Lecture "Understanding the Human Mind Through Birds" 2019/5/12

To date, with researchers from the Center for Human-Intelligence Research—established based on the comprehensive agreement between Keio University and RIKEN—playing a central role, Keio University has been recognized for undertaking important research as a member institution in the "Project for Elucidating the Entirety of Brain Function Networks through Innovative Technologies" by the Japan Agency for Medical Research and Development (AMED), which started in FY2014. As a result, a multi-beam scanning electron microscope with 61 beams for the world's fastest wide-area imaging—one of only three such units in regular operation worldwide—was installed at Keio University in 2016. A state-of-the-art, super-resolution fluorescence microscope equipped with 64 detectors was installed at Keio University in FY2017, and multiple achievements utilizing these instruments are now being reported. Furthermore, to carry out the project item on cell-type-specific long-term imaging of the marmoset brain using fluorescent calcium imaging technology, the newly developed two-gram ultra-compact fluorescence microscope, nVista, and the nVoke microscope, which also enables optical stimulation, were introduced in 2017 and 2018, respectively. An upgrade including an autofocus function was implemented in February 2019. This has made it possible to visualize neural activity in the living primate brain with fluorescent probes and to artificially induce neural activity through optical stimulation, with the results now being prepared for publication.