京都大学教育研究振興財団助成事業
成 果 報 告 書
２０２４ 年 ５ 月 ７ 日
公益財団法人京都大学教育研究振興財団
会 長

藤

洋 作

様

所属部局

京都大学iPS細胞研究所（CiRA）

職

名

教授

氏

名

齊藤 博英 （サイトウ ヒロヒデ）

令和 ４ 年度

助 成 の 種 類

・国際会議開催助成

国 際 会 議 名

iPSZürich 2024 Symposium & CiRA-IREM Joint Workshop

開 催 期 間2024 年 4 月 4 日

開 催 場 所

チューリッヒ大学 （スイス）～ 2024 年 4 月 5 日

者

内訳：修士学生（28名）、博士学生（44名）、研究員（29名）、グループリー
ダー（29名）、企業員（30名）、その他（23名）
男性（105名）、女性（74名）、言いたくない（4名）
チューリッヒ大学（69名）、京都大学（12名）、その他（102名）

成 果 の 概 要

タイトルは「成果の概要／報告者名」として、A４版２０００字程度・和文で作成し、添付して下さい。 「成果の概要」以外に添付する資料 □ 無☑ 有(プログラム)

総数：１８３名
参加事 業 に 要 し た 経 費 総額8,620,000円

うち当財団からの助成額

1,000,000円

その他の資金の出所

SNSF, MaxWell, 3Brain, Bucher, Seed, IotaScience, Stemcell Technologies, OLS,
Agilent, Miltenyi, Univeristy of Zurich, iPS細胞研究所, 京都大学教育研究振興財団

経 費 の 内 訳 と 助 成 金 の 使 途 に つ い て
費会 計 報 告当財団の助成に
つ
い
て

目

金

額

（円）

雑費（消耗品含）
会議費（通信費含む）
交際費
旅費交通費
謝金（現地協力者）
諸会費

170,000
1,190,000
4,182,000
2,550,000
272,000
256,000

合8,620,000

計

財団助成充当額 （円）50,00050,000900,000

1,000,000

今回はご支援いただき、心より感謝申し上げます。
おかげ様で、再生医療に関する共同研究及び若手研究者の頭脳循環を推進するための共同ワークショップを開催することができました。
今後も本助成を継続していただければ幸いに存じます。

【成果の概要 / 京都大学iPS細胞研究所 教授 齊藤 博英】
京都⼤学 iPS 細胞研究所（CiRA）とチューリッヒ⼤学の再⽣医療研究所（Institute of Regenerative Medicine, IREM）は再⽣医療を世界に広めるという共通の⽬標のもと、2017 年から共同研究を⾏っています。IREM は 2022 年より毎年、iPS細胞の医療応⽤に向けた研究の進展にスポットライトを当てた「iPSZürich シンポジウム」を主催しています。今年度は、iPSZürich シンポジウムと併せて CiRA-IREM合同ワークショップを開催し、両研究所の研究を紹介するとともに、両機関の⼤学院⽣や若⼿研究者が将来、研究活動のために海外に⾏くことを奨励するプログラムを企画しました。
京都⼤学教育研究振興財団、スイス科学財団(SNSF)、他のアカデミアや企業のスポンサーからの助成により、iPSZürich シンポジウム 2024 と CiRA-IREM 合同ワー
クショップは 2024 年 4 ⽉ 4 ⽇〜5 ⽇に 180 名以上の登録参加者（現地、オンライ
ン併せて）が参加し、スイスのチューリッヒ⼤学で無事に開催されました。
CiRA を代表して今回のシンポジウムと合同ワークショップに参加する⼤学院⽣
や若⼿研究者を⽀援するため、5 つのトラベルアワードを設けることができました。スイス到着の当⽇（4 ⽉ 3 ⽇）、⼀同は IREM を訪問し、両研究所の教授が次世
代再⽣医療を世界に広めるという共通の⽬標についての意⾒交換会を⾏いました。
ディスカッション後、CiRA の教員とトラベルアワード受賞者は、IREM の研究室と患者検査センターの⾒学を⾏いました。最後に、IREM の若⼿研究者や学院⽣が起業家精神を育み、科学的なアイデアをビジネスとして成功させるために、IREM メン
バーが利⽤参加できるバイオメッド・アントレプレナーシップ・プログラムの紹介を受けました。
翌⽇（4 ⽉ 4 ⽇）、チューリッヒ⼤学のシティキャンパスで CiRA-IREM 合同ワー
クショップが開催されました。Jessica Plucain 先⽣と Kelvin Hui 先⽣による両⼤学や研究所の簡単な紹介をしました。それから、京都⼤学⽣あるいは京都⼤学での勤務経験があり、その後ヨーロッパに移った 4 名の招待講演者によるパネルディスカ
ッションが⾏われ、⾃⾝の海外での研究経験を伝えることで、参加者の海外留学への挑戦を推奨しました。パネルディスカッションの後に、招待講演者、CiRA-IREM
トラベルアワード受賞者、参加者のより活発な交流を促すため、コーヒーブレイク
交流会が⾏われました。最後は、Leslie Reinhardt 先⽣と Kelvin Hui 先⽣がチューリッヒ⼤学や CiRA ⼜は京都⼤学での短期及び⻑期の研究滞在の助成⾦制度をワーク
ショップ参加者に説明しました。⼣⽅には、IREM 所⻑ Simon P. Hoerstrup 先⽣とCiRA 所⻑髙橋淳先⽣による合同公開講演会が開催され、スイスと⽇本における再⽣医療の現状について議論が交わされました。

1

最終⽇（4 ⽉ 5 ⽇）に、iPSZürich シンポジウム 2024 が開催されました。CiRA の主任研究者である Knut Woltjen 先⽣と⿑藤博英先⽣の 2 ⼈がシンポジウムの「Development and Aging」と「Gene Regulation」のセッションで講演しました。さらに、「Modelling in Health and Disease」と「Translational Application」のセッショ
ンで、Laurine Cabon 先⽣（Roche Institute of Human Biology）と Ute Modlich 先⽣（IREM）の招待講演が⾏われました。CiRA-IREM トラベルアワード受賞者を含む 12名の講演も⾏われました。また、シンポジウムでは 2 つのポスターセッション（計40 ポスター）も⾏われ、CiRA-IREM トラベルアワード受賞者が CiRA での研究業績を発表しました。

CiRA-IREM トラベルアワード受賞者：網本 直弥（⼤学院⽣）、中村 英美⾥（⼤学院⽣）、Linh Nguyen（⼤学院⽣）、⼭川達也（研究員）、Zujur Denise（研究員）

参加者の内訳：
修⼠学⽣（28 名）、博⼠学⽣（44 名）、研究員（29 名）、グループリーダー（29名）、企業員（30 名）、その他（23 名）
男性（105 名）、⼥性（74 名）、⾔いたくない（4 名）
チューリッヒ⼤学（69 名）、京都⼤学（12 名）、その他（102 名）

2

CiRA の教員とトラベルアワード受賞者 IREM に訪問

iPS 細胞研究所／京都⼤学で研究経験のパネルディスカッション

3

CiRA-IREM トラベルアワード受賞者の⾃⼰紹介

CiRA と IREM の若⼿研究者の交流

4

iPS 細胞研究所所⻑髙橋淳先⽣の合同公開講演会

5

Welcome to the
iPSZurich Symposium 2024
4-5th April 2024 • Zurich, Switzerland

Workshop: KOL-F-101
Public Lecture & Symposium: KOH-B-10
Rämistrasse 71
8006, Zurich1

In recognition that this event was made possible through our kind sponsors,please read about them below and vist their booths.

ors 2024

Please don’t print me.

2

3

Voting QR codes
Don’t forget to vote for the your favourite poster and presentation using the
QR-codes below.

Favourite Poster

https://strawpoll.com/eJnvvakQ4nv

Favourite Oral Presentation

https://strawpoll.com/wAg3AoL7My8

4

Sponsors
Swiss National Science Foundation

More info here

The Swiss National Science Foundation (SNSF) funds excellent research at universities and otherinstitutions – from chemistry to medicine to sociology. Thousands of teams are generatingknowledge for a better future for all. Together with our partners, we play a key role in shaping
research in Switzerland.
The Kyoto University Foundation

KYOTO UNIVERSITY
FOUNDATION

More info here

The Kyoto University Foundation for Education and Research provides grants for the promotionof high-level education and research at Kyoto University, the continuation and development ofimportant academic fields that are not necessarily in great demand from a social and economic
perspective, human resource development for young researchers, and contribution activities forthe revitalization of local communities, thereby contributing to the promotion of science,technology, and other interests.
UZH Alumni

More info here

Through alumni organizations, former students not only stay connected to each other and theiralma mater, but also actively shape the latter’s vitality. They thus remain an essential part of the
UZH family even after graduation. The UZH alumni networks boasts 18’400 members, across 25
alumni organisations.
3BrainMore info here

3Brain is the world’s first company to connect cells with sophisticated silicon chips in cell culture
plates. Stemming from their passion for technology and scientific progress, their co-founders
have worked for more than 15 years on CMOS-powered cell-electronic interfaces with the aimof boosting research in major fields like neuroscience, ophthalmology and cardiology.
MaxWell Biosystems

More info here

MaxWell Biosystems AG is an electronics and biotechnology company based in Zurich,
Switzerland that provides instrumentation and solutions to advance neuroscience research andaccelerate drug discovery. The company was incorporated in September 2016. Applications
include a electrophysiology cell-imaging platform and High-Resolution MEA Technology basedon micro-sensor fabricated in CMOS technology.
Bucher Biotec AG

More info here

Bucher Biotec AG is a Swiss distributor representing leading manufacturers of highly innovative
life science research instrumentation. We are extremely proud of our distinguished customer
base in the pharmaceutical, biotechnology, agricultural, food and related industries, in all life
science research oriented academic institutions, in numerous governmental, clinical andenvironmental labs and in all university hospitals. Our highly competent, well educated team is
focused on understanding our customer’s needs in order to be able to propose optimal solutions
for the demanding research tasks.
NanoCellect Biomedical, Inc.

More info here

NanoCellect is committed to empowering every scientist to make discoveries one cell at a time,with modern and simple technologies for to advance cell based assays that are affordable,compact, and easy-to-use. Our microfluidic flow cytometry platforms enable biomedical
scientists to analyze and sort cells required for drug discovery, single cell-omics, cloning, and basic
research. The company was founded in late 2009 as a spinout from UCSD and dedicated 6 years
developing the foundation of the WOLF’s technology before introducing the WOLF to early
adopters in 2016. Initial funding of R&D was graciously provided by multiple NIH SBIR grants and5contracts. Additionally, we are backed by Illumina Ventures, FusionX Ventures, Anzu Partners,
Agilent Technologies, Vertical Ventures and other private investors.
SEED Biosciences

More info here

SEED Biosciences developed DISPENCELL with the vision of democratising the use of single cells
in research. DISPENCELL enables scientists to isolate single cells three times faster and at a cost
10 times lower than existing solutions. Its ambition is to set new standards in single-cell biology
in order to accelerate the translation of precision medicine from research to personalised
therapies for the benefit of patients.
Miltenyi Biotec

More info here

For more than 30 years, Miltenyi Biotec has played an important role in the design, development,
manufacture, and integration of products that empower the advancement of biomedical
research and enable cell and gene therapy. Miltenyi has 17,000 Products available worldwide
and at the heart of their business lies MACS Technology, a magnetic cell separation method
based on the use of MACS Columns and MACS MicroBeads.
OMNI Life Science

More info here

OMNI Life Science is your Partner for Solutions in 3D Cell Culture, Cell Counting, Cell Assays, Cell
Imaging and Microbiology. Innovative systems for research in Life Sciences: From cell culture to
real-time cell assays, cell counting and microbiology. 3 Reasons to partner with OLS: (1) matchyour need with the broad range of excellent cell analysis instruments, (2) Gain best results in lesstime with user-friendly instruments and powerful software, (3) Benefit from the OLS experts´know-how, instrument trainings and ongoing service.
Agilent

More info here

Agilent is a global leader in life sciences, diagnostics and applied markets, recognized for
uncompromising integrity in all we do. Our mission is to advance quality of life by focusing our
expertise in six key markets. With a team of approximately 18,000 employees spread across theworld, our global footprint includes sales offices, logistics centers, and business facilities in threemajor geographies.
iotaSciences

More info here

We are a young and dynamically growing spin-out company from Oxford, UK. The company was
founded in summer 2016 by scientists from the Department of Engineering Science and the Sir
William Dunn School of Pathology, University of Oxford, UK. Since its inception, iotaSciences has
been committed to offer cell researchers highly user-friendly, reliable and powerful technologies
with unique capabilities that greatly simplify and facilitate breakthrough advances in cell biology.
STEMCELL Technologies

More info here

STEMCELL Technologies provides high-quality cell culture media, cell separation technologies,
instruments, accessory products, and educational resources to scientists around the world
working on stem cell, immunology, cancer, regenerative medicine, and cellular therapy research.
At STEMCELL, we are dedicated to improving lives through advanced knowledge and scientificdiscovery, through our commitment to fostering diversity and inclusion in STEM and the life
sciences industry, and through our investments in sustainability, community, and social
responsibility.

6

4th April 2024 – CiRA-IREM Joint Workshop
07:30 – 08:00

Registration

Registration desk

08:00 – 08:30

Welcome & Introduction KU and UZHRoom KOL-F-101

Welcome and Introduction of Kyoto University and University of Zurich as a hub of life
science and innovations
08:30 – 09:30

Experience Reports

Room KOL-F-101

iPSZürich Workshop
April 4 , 2024 • Zurich, Switzerland
Experience Reports of Swiss living/studying in Japan and vice versa
Invited speakers: Thomas Maurissen (Industry), Reiko Akiyama (UZH),thSora Matsumoto, (EMBL), Debora Kehl (Industry)

09:30 – 10:30

April

Lichthof, Floor D

Meet & Greet
Meet and greet with our invited workshop guests with Gipfeli andcoffeeReports of Scientific Exchanges Room KOL-F-101
10:30 – 11:00 Experience
Funding opportunities
Leslie Reinhard,
Globalfor
Affairs,
UZH
Funding
SourceResearch
Stays Abroad
Dr. Kelvin Hui, CiRA

18:00 – 19:30

Public Lecture

Public Lecture

Room KOH-B-10

Prof. Hirohid
Kyoto Univ

Public lecturethby Prof. Jun Takahashi, CiRA and Prof. Simon P. Hoerstrup, IREM

April 4 , 2024 • 6PM • UZH, Rämistrasse 71

REG
S

«

Institute for Regenerative
Medicine • IREM

Prof. Jun Takahashi
Kyoto University«De

Prof. Simon P. Hoerstrup
University of Zurich«The Current State of Regenerative Medicine
in Japan and Switzerland»

«Tran
7

5th April 2024 – Symposium
08:00 – 08:30

Welcome & Registration

Registration desk

08:30 – 10:00

Session 1: Development and Aging

Room KOH-B-10

Guest speaker: Prof. Knut Woltjen (30 +10 min), CiRA
Abstract Selected Speaker (10+3 min):
Linh Nguyen, Kyoto University, Japan
Ermanno Malagola, University of Zurich, Switzerland
Ramon Pfaendler, ETH Zurich, Switzerland
10:00 – 10:30

Coffee Break

Lichthof, Floor D

Refreshments provided in the Lichthof
10:30 – 12:00

Session 2: Modelling in Health and Disease

Room KOH-B-10

Guest speaker: Dr. Lauriane Cabon (30 +10 min), Roche
Abstract Selected Speaker (10+3 min):
Edna Grünblatt, University of Zurich, Switzerland
Sarah Guimbal, University of Bern, Switzerland
Martina Nubie, University of Zurich, Switzerland
Gold sponsor: MaxWell Biosystems (5 min)
12:00 – 13:00

Lunch

Lichthof, Floor D

Refreshments provided in the Lichthof
13:00 – 14:00

Poster Session I

Lichthof, Floor D

Selected posters session (Odd numbers must be by poster)
14:00 – 15:30

Session 3: Gene Regulation

Room KOH-B-10

Guest speaker: Prof. Hirohide Saito (30 +10 min), CiRA
Abstract Selected Speaker (10+3 min):Alberto Catanese, Ulm University, Germany
Tatsuya Yamakawa, Kyoto University, Japan
Jiang-An Yin, University of Zurich, Switzerland
15:30 – 16:30

Poster Session II

Lichthof, Floor D

Selected posters session (Even numbers must be by poster)8

16:30 – 18:00

Session 4: Translational Application

Room KOH-B-10

Guest speaker: PD Dr. Ute Modlich (30 +10 min), IREM, UZH
Abstract Selected Speaker (10+3 min):Emiri Nakamura, Kyoto University, Japan
Lewis Jones, ETH Zurich, Switzerland
Blandine Clément, ETH Zurich, Switzerland
Gold sponsor: 3Brain (5 min)
18:00 – 18:30

Closing remarks and Awards

Room KOH-B-10

18:30 – 20:00

Networking & Apéro

Lichthof, Floor D

iPSZürich organizing team

Melanie Generali

Vanessa Budny

Melanie Eschment

Kelvin Hui

Clara DuréDavid Taborsky

9Experience Reports of Scientific Exchanges
Funding Source for Research Stays Abroad

Public Lecture

Prof. H
Kyoto

PublicApril 4th, 2024
• 6PMLecture• UZH, Rämistrasse 71

RE
Institute for Regenerative
Medicine • IREM«

Prof. Simon P. Hoerstrup
University of ZurichProf. Jun Takahashi
Kyoto University

«T

«The
Current
State ofMedicine
Regenerative
The
Current
State of Regenerativein Japan andMedicine
Switzerland
in Japan and Switzerland»"CIRA'S MISSION IS NOW IN PROGRESS"The Center for iPS Cell Research and Application was established in 2010 with a clear mission:
the medical application of iPS cells and related technologies. We now have three researchbuildings with about 600 faculty members, researchers, and students in Kyoto, a beautiful
historical city in Japan. As you may know, iPS cells have two characteristics: self-renewal andpluripotency, meaning they can proliferate indefinitely as stem cells, and under appropriate
conditions, they can differentiate into the main cell types of our body. Taking advantage ofthese characteristics, we are trying to apply iPS cells for regenerative medicine, namelytransplanting iPS cell-derived somatic cells to replace lost or damaged cells. Alternatively, we
use iPS cells to generate disease models to investigate disease mechanisms and develop new
drugs. In my talk, I will introduce our efforts and results in both directions, especially mywork on an iPS cell-based therapy for Parkinson's disease.
“IREM – A BLUEPRINT FOR TRANSLATIONAL REGENERATIVE MEDICINE”
Based on bio-inspired principles and developmental engineering/re-engineering
approaches, the Institute for Regenerative Medicine (IREM) aims to replace or regenerate
dysfunctional human cells, tissues, or organs with the goal of restoring normal functions. By
using its designated clinical translational facilities, ranging from Cleanroom Technologies(Center for Therapy Development), iPS Core Unit, and a Clinical Trial Unit (Center for
Prevention and Dementia Therapy), IREM fulfills its mission to translate excellent basic
research into innovative clinical application and advancing molecular life sciences into the
future.
Furthermore, the interaction with Wyss Zurich, a unique center with the ETH Zurich and the
University of Zurich, drives the fields of regenerative medicine and robotics, together withbionics technologies. Wyss Zurich brings together some of the world’s leading experts toform multidisciplinary teams, pooling their knowledge and expertise.
10Invited Workshop Speakers
Maurissen, ThomasThomas obtained his BSc/MSc in Bioengineering at EPFL, during which he participatedin a summer exchange program between EPFL and the University of Tokyo. This
experience in Japan sparked his interest to apply and later obtain his PhD from Kyoto
University, Graduate School of Medicine. Thomas greatly enjoyed 3.5 years of iPS cellresearch at CiRA in the Woltjen lab. Now, he is a scientist and research project leader
in a pharmaceutical company, working on retinal/vascular diseases.

Akiyama, Reiko

Reiko is a plant ecologist by training and is a lab manager at the Department of
Evolutionary Biology and Environmental Studies at the University of Zurich. During her
MSc study at Kyoto University, she stayed at Uppsala University in Sweden as an
exchange student. With a PhD from Uppsala University, she came to Switzerland as a
postdoc and worked with collaborative research projects between Switzerland andJapan. She embraces the culture and nature of both countries.

Matsumoto, Sora

Sora is currently pursuing a PhD in Dr. Sinem Saka's group at the European Molecular
Biology Laboratory (EMBL) in Heidelberg. Prior to this, he earned his MSc and BSc at
Kyoto University under the guidance of Dr. Hirohide Saito in CiRA. Through acollaborative project with researchers in France, he had the opportunity to familiarize
himself with the European research environment, which eventually led him to apply for
his PhD studies at EMBL. His research interests span across RNA biology, synthetic
biology, liquid-liquid phase separation, and spatial omics. Outside the lab, Sora enjoys
bouldering and cooking Japanese food.

Kehl, Debora

Debora Kehl holds a PhD in Integrative Molecular Medicine from the University of
Zurich and currently works in clinical research in the pharmaceutical industry in
Switzerland. As a postdoctoral fellow at the Institute for Regenerative Medicine (IREM),
Debora supported the establishment of a scientific collaboration with the Center for
iPS Cell Research and Application (CiRA) in Kyoto. As visiting researcher she stayed at
CiRA and focused on iPSC-derived cardiomyocytes to regenerate ischemic heart tissue.

iPSZürich Symposium 2024 CiRA-IREM Travel Award Recipients

DeniseZujur

Linh
NguyenEmiri
Nakamura

Naoya
AmimotoTatsuya
Yamakawa

11

Invited Symposium Speakers
Woltjen, KnutMore info here

GENOME AND EPIGENOME EDITING TOOLS FOR STEM CELL RESEARCH
Establishing control over the genome and epigenome is desirable for the
study of human health and aging as well as the development of reliable
cellular therapeutics. To achieve accurate disease models, we havefocused on predicting DNA repair outcomes following gene editing inhuman induced pluripotent stem (iPS) cells with CRISPR-Cas9. In this way
we have established an isogenic allelic series for single nucleotide
variants, provided a genome-wide resource for creating deletion
mutations, and are exploring the design requirements for cutting edge
methods such as Prime Editing. In an alternative strategy, we are
employing epigenome editing for cell engineering without permanent
genetic modification. Examples of partial reprogramming of somatic cells
for biological age reversal and targeted gene repression for the creationof ‘universal’ iPS cells will be presented. The current challenges of using
epigenome editing for cellular rejuvenation and cell therapies will be
discussed.
Saito, Hirohide

More info here

RNA SYNTHETIC BIOLOGY TO PROGRAM CELLS
Research in synthetic biology, which aims to understand biologicalphenomena by creating biomolecules and life systems, is advancing andshows promise in fostering new technological breakthroughs. In thispresentation, I will introduce studies based on the interactions between
RNA and RNA-Protein (RNP). Our team has been progressing research onmRNA switches that can regulate translation based on the state of thecell. We have developed a switch that turns off translation in response
to proteins that characterize cells and microRNA (miRNA) - referred to as
the miRNA-responsive OFF switch. However, with just this OFF switch,
purification efficiency was a challenge for certain cell types. Recently, we
have developed an innovative switch that, contrary to the OFF switch,
activates translation in response to miRNA, known as the miRNAresponsive ON switch. In the development of this ON switch, we
discovered a unique RNA design technique where an artificial sequenceis inserted downstream of the mRNA’s poly-A tail. By combining both the
ON and OFF switches, we successfully purified cardiomyocytes
differentiated from iPS cells at high efficiency and eliminated unwanted
cells without the need for equipment like cell sorters. Along with these12RNA switch technologies, this talk will also introduce the latest
achievements in cell reprogramming studies based on RNA.
Modlich, Ute

More info here

GENE THERAPY FOR INHERITED BLOOD DISORDERS
There are more than 7000 inherited disorders caused by monogenetic
mutations, many of which affect the blood and immune system. Because
all blood cells are produced by hematopoietic stem cells (HSC),
transplantation of allogeneic HSC from healthy and compatible donors
can cure these disorders. However, suitable donors are not always
available (e.g. in 2/3 of Caucasian patients). In these cases, gene therapy
using autologous patient HSC is a viable alternative, using an ex vivo genemodification, either adding a healthy gene copy or correcting themutated gene, and transplantation back into the patients.
In our research group, we focus on gene therapy affecting myeloid cells,
especially macrophages, or megakaryocytes and platelets. For targeted
expression, we developed a lentiviral vector platform for expression inthese cell types. Due to limited access to patient-derived primary cells,
the use of iPSC-derived hematopoietic cells represents an important test
platform for developing gene therapies. Using iPSC with disease-specificmutations, the specificity of transgene expression and correction of thephenotype can be studied. Differentiation of hematopoietic cells fromiPSC is guided by specific transcription factors, typically induced bycytokines. By inducible overexpression of these transcription factors
during hematopoietic commitment, blood cell differentiation can besupported. Inducible expression systems in iPSC for megakaryocytic
development and the challenges of tight expression control will be
discussed.
Cabon, Lauriane

More info here

INSTITUTE OF HUMAN BIOLOGY AT ROCHE
In this talk, Lauriane will present how novel therapeutic modalities in the
oncology space led to the establishment, improvement and adoption of
adult stem cell derived organoids assays in drug development. T-cellbispecific (TCBs) antibodies are an innovative tool for harnessing theimmune system to fight cancer whereby TCBs simultaneously bind to a
T-cell and a tumor antigen resulting in tumor killing. TCBs efficacy reliesin part on the ability of the T-cell to access the tumor tissue; while theirsafety is driven by levels of expression of the tumor antigen in normal
tissues. Relying on human relevant preclinical assays to evaluate andbetter understand the benefit/risk ratio of such molecules became
crucial over the past years. Lauriane will more specifically highlight the13

advances her team and colleagues at Roche have made by introducing
lymphocytes into organoids assays, from peripheral blood mononuclear
cells to tissue resident lymphocytes. She will then outline a few otherexamples of stem cell derived organoids assays applied in theophthalmology and immunology disease areas.

Selected Speaker Abstracts
Session 1: Development and Aging
Nguyen, Linh

Kyoto University, JP“ALL MEN ARE CREATED EQUAL" BUT ARE ALL IMSCS CREATED EQUAL?
Mesenchymal stem cells (MSCs) are a promising cell source in regenerative medicine fortreating various diseases owing to their immuno-regulatory effects and multi-lineage
differentiation capacity. The defining characteristics of MSCs are 1) adherence to plastic
culture dishes, 2) expression of specific surface markers CD73, CD90, and C105, and 3)
differentiation into osteoblasts, chondrocytes, and adipocytes. However, MSCs extracted
from the human body often show inconsistent quality and heterogeneity. Although iPSCderived MSCs are expected to overcome such problems, the nature of iPSC-derived MSCs
from different lineages remains unknown, and comprehended research is required. In thisstudy, we successfully generated MSCs from the same human induced pluripotent stem cells(iPSCs) through different lineage-specific routes, including cranial neural crest cells, trunk
neural crest cells, paraxial mesoderm (somite), lateral mesoderm, and limb mesenchyme.
We then compared these differentially originated iPSC-derived MSCs with tissue-derived
MSCs from adipose tissue, bone marrow, muscle, and dental pulp. All five types of iPSCderived mesenchymal stem cells fulfilled the criteria but showed different capacities for
multi-lineage differentiation, depending on the differentiation route. Specifically, somite,cranial neural crest, and limb mesenchyme-derived MSCs showed higher osteogenic potency
than trunk neural crest and lateral plate mesoderm-derived MSCs, implying the effect ofdevelopmental origin on different iMSCs’ characteristics. PCA plot distinctly illustrates thevariations in global gene expression patterns among the iMSCs. Furthermore, dental pulpderived MSCs and neural crest-derived MSCs are shown not to cluster together, suggestingthat factors beyond cellular origins, such as functional attributes or environmental
influences, might play a role in determining the global gene expression of MSCs. Our research
represents a thorough investigation into different iMSCs’ characteristics, contributing to agreater understanding of the heterogeneity associated with iPSC-derived MSCs, which willbe critical for their therapeutic applications.

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Malagola, Ermanno

University of Zurich, CH

ISTHMUS PROGENITOR CELLS CONTRIBUTE TO HOMEOSTATIC CELLULAR TURNOVER AND
SUPPORT REGENERATION FOLLOWING INTESTINAL INJURY (MOUSE)The currently accepted intestinal epithelial cell organization model proposes that crypt basecolumnar (CBC) cells marked by high levels of Lgr5 expression represent the sole intestinal
stem cell (ISC) compartment. However, previous studies have indicated that Lgr5+ cells are
dispensable for intestinal regeneration, leading to two major hypotheses: one favoring thepresence of a quiescent reserve stem cell population, the other calling for differentiated cell
plasticity. To investigate these possibilities, we studied crypt epithelial cell organization,
during homeostasis and regeneration, in unbiased fashion, via high-resolution single-cell
profiling. These studies, combined with in vivo lineage tracing, show that Lgr5 is not a specific
ISC marker and that stemness potential exists beyond the crypt base in the isthmus region,
whose cells, contrary to differentiated cells, participate in tissue homeostasis and support
intestinal regeneration. Our results provide a novel model of organization for the intestinal
crypt epithelium in which stemness potential is not restricted to CBC cells and suggestingthat neither de-differentiation nor reserve stem cell populations are drivers of intestinal
regeneration.
Pfaendler, Ramon

ETH Zurich, CH

A SYSTEMS VIEW OF CELLULAR STATE HETEROGENEITY IN HUMAN PLURIPOTENT STEM CELLS
Early human embryogenesis relies on cellular fate transitions of pluripotent stem cells
orchestrated by various signalling pathways and spatio-temporal dynamics. While recent
progress deepened our understanding of the gene regulatory programs underlying theseprocesses, the molecular determinants governing cellular state heterogeneity of pluripotent
stem cells at the single-cell level remain incompletely quantified and understood. Here, weinvestigate the functional and phenotypic variability of induced pluripotent stem cells (iPSCs)
upon drug perturbations using a combination of high-content microscopy, computer vision,and high-throughput molecular profiling. Quantification of the high-content imaging datausing self-supervised deep learning unveiled diverse single-cell morphological states of iPSCs
across drug mode-of-actions. In addition, multiplexed transcriptomics and high-throughput
proteomics revealed condition-specific expression dynamics of key proteins and transcripts
involved in early differentiation. Intriguingly, integrating single-cell morphological stateswith this multimodal molecular data enables us to link the underlying molecular processes
that govern iPSC cellular states. Globally, this suggests that iPSCs navigate an intricate path
between simultaneously changing cellular morphology and fate. Overall, our study provides
valuable insights into the molecular landscape underlying phenotypic heterogeneity in acellular system characterised by maximal developmental plasticity across scales.

15Session 2: Disease Model

Grünblatt, Edna

University of Zurich, CH

ADHD NEURODEVELOPMENTAL ALTERATION ASSOCIATED TO WNT PATHWAYS, OXIDATIVESTRESS AND INFLAMMATION: PERSONALIZED TREATMENT PROSPECTS
Due to its polygenic and phenotypic spectrum, attention-deficit hyperactivity disorder(ADHD) is a complex neurodevelopmental disorder. Over 5% of children and adolescentsworldwide are affected, with 60% persisting into adulthood. While the cause of ADHD is
unknown, neurodevelopmental delays associated with Wnt-pathways and subsequentoxidative stress (OS) and inflammation suggest plausible causes. Response to
Methylphenidate (MPH), a psychostimulant, is one of the highest in mental disorders, yetsome do not benefit due to either partial or no response. Nevertheless, the mechanism ofaction remains unclear. Some patients also benefit from polyunsaturated fatty acids (PUFAs),
particularly omega-3, the effect that may be attributed to its anti-inflammatory and anti-OS
properties. We used patient-derived induced pluripotent stem cells (iPSCs), neural stem cells(NSCs), and forebrain cortical neurons (FCNs) to model ADHD to identify the critical time
point for alterations and whether we could identify treatment-specific response. Intriguingly,
modifications began only at the NSC phase, where cells establish neural fate. Compared tocontrols, ADHD cells exhibited decreased growth-rates, linked to alterations in Wnt-activity.
These alterations associated with personalized genetic predisposition and clinical scores,
enhancing the model's clinical translation ability. Additionally, ADHD FCNs showed elevated
OS and inflammatory markers, supporting clinical findings. MPH and omega-3 reduced
cellular alterations, which was encouraging. Notably the response to MPH was only detected
in clinically responsive patients, suggesting specific mechanisms. Overall, our findings
contribute to explaining ADHD's etiology and treatment mechanisms in patient-specificneural cell models, paving the path to novel approaches, preventive measures, anddiscoveries of new therapeutic targets.
Nubie, Martina

University of Zurich, CH

DEVELOPMENT OF HUMAN IPSC-BASED MODELS TO TEST MICROGLIA-DIRECTED GENE
THERAPY FOR NEURODEGENERATION
The deficiency of progranulin (PGRN), caused by mutations in the granulin (GRN) gene, is
responsible for two fatal neurodegenerative diseases: neuronal ceroid lipofuscinosis-11(CLN11) in children and young adults, and frontotemporal dementia (FTD) with typical onset
before 65 years of age. Currently, no disease-modifying treatment is available. We develop
an ex vivo hematopoietic stem and progenitor cell (HSPC) gene therapy that targets
microglia, the brain-resident immune cells critically involved in the pathology. In an
autologous transplantation setting, the gene-modified patient HSPCs generate new
microglia-like progeny that secrete higher levels of PGRN. For this gene therapy, we designed
a lentiviral vector (LVV) that drives phagocyte-specific PGRN expression. Prior to in vivo
application, we tested the LVV in a model of microglia differentiated from induced
pluripotent stem cells (iPSCs) carrying mutations in the GRN gene. We successfully generated
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microglia expressing typical markers such as IBA1, TMEM119 and SALL1. Upon iPSC
transduction with the gene therapy LVV, we could increase PGRN expression in the microglia
to physiological levels. We also investigated the dynamics of transgene expression driven by
our myelospecific promoter by monitoring the emergence of lineage-restricted myeloid
markers during differentiation. We confirmed that our LVV is inactive in early differentiation
and becomes activated during myeloid specification. To mimic the integration of PGRNenhanced microglia in the patient’s brain, other brain cell types need to be included in thismodel. In a pilot experiment, we defined the culture conditions that support thedifferentiation of iPSC-derived neurons, astrocytes and microglia. The development of thisco-culture model will allow not only to test candidate LVVs for microglia gene therapy, butalso to investigate the underlying human-specific pathogenic mechanisms of GRN-related
neurodegeneration.
Guimbal, Sarah

University of Bern, CH

NEW MOLECULAR UNDERPINNINGS OF BBB DYSFUNCTION IN MULTIPLE SCLEROSISIntroduction: Blood-brain barrier (BBB) breakdown is amongst the earliest pathologicalhallmarks in multiple sclerosis (MS). The mechanisms leading to BBB dysfunction are
incompletely understood and are generally thought to be a consequence of the autoimmune
neuroinflammatory process in MS. Objective: We challenge this view and ask if intrinsic
alterations in BBB endothelial cells manifested at the genetic, epigenetic, transcriptional, andphenotypic level contribute to altered BBB function. Methods: We made use of humaninduced pluripotent stem cells (hiPSCs) derived from 6 healthy controls (HC) and 10 MS
patients and differentiated them into brain microvascular endothelial cell (BMEC)-like cells
as in vitro model of the BBB. We performed a transcriptomic analysis on the donors available(3 HC and 4 MS patients) via RNA sequencing on HC and MS-derived BMEC-like cells
stimulated with TNF-α and IFN-γ and unstimulated. Results: The RNA sequencing analysis
showed an increase of regulated genes in unstimulated BMEC-like cells compared to thestimulated condition, strengthening that BBB may contribute directly to MS pathology.
Moreover, it also revealed a strong modulation of the Semaphorin-4D (SEMA4D) signalling
pathway in unstimulated MS-derived BMEC-like cells compared to the controls. We
confirmed, via western blot and immuno-staining, the expression of SEMA4D and its
downstream effectors, RHOB and ROCK2. EECM-BMEC-like cells were treated with arecombinant protein for SEMA4D and showed decreased mRNA expression of SEMA4D anda cytoskeletal reorganization. Conclusion: Our study suggests that SEMA4D and its
downstream effectors could play a role in BBB dysfunction in the context of MS.

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Session 3: Gene Regulation
Catanese, Alberto

Ulm University, GER

HETEROZYGOUS KNOCKOUT OF SYNAPTOTAGMIN13 PHENOCOPIES ALS FEATURES IN
HUMAN MOTOR NEURONS
Synaptic alterations represent a common feature shared across the heterogenous
pathogenetic spectrum of Amyotrophic Lateral Sclerosis (ALS). In particular, we haverecently shown that the presynaptic SNARE machinery is detrimentally impaired in ALS
motoneurons (MNs), which indeed display reduced firing properties. In this work, we aimed
at identifying specific presynaptic targets actively contributing to neuronal vulnerability anddisease progression in ALS. We combined multi-omics and deep learning strategies withhuman iPSC-derived MNs to highlight Synaptotagmin 13 (Syt13) as a candidate synaptic
protein contributing to neuronal vulnerability in ALS. By using CRISPR-Cas9 technology, wethen investigated whether the heterozygous loss of Syt13 is sufficient to resemble an ALS
phenotype. Syt13+/- hiPSC-derived MNs display a progressive manifestation of typical ALS
hallmarks such as loss of synaptic contacts and accumulation of aberrant aggregates. By
comparing the transcriptome of Syt13-deicificent cells to Syt13+/+ ones, we found asignificant impairment in biological mechanisms involved in motoneuron specification andspinal cord differentiation. In addition, we identified an astonishing correlation of theSyt13+/- transcriptome with an ALS fingerprint generated using RNAseq data from human
MNs and post mortem spinal cord samples. This significant overlap converged toward a
detrimental upregulation of neuronal death and pro-inflammatory response, which was
linked to a dysfunctional Akt-GSK axis. Our data show for the first time that the heterozygous
loss of a single member of the synaptotagmin family is sufficient to trigger a typical ALS
phenotype leading to the death of human MNs, thus revealing novel insights into theselective vulnerability of this cell population.
Yamakawa, Tatsuya
NOVEL PATHOGENESIS OF DIAMOND-BLACKFANREGULATION OF PROTEOSTASIS

Kyoto University, JP
ANEMIAEXPLORED

THROUGH

Diamond-Blackfan anemia (DBA) is a congenital bone marrow failure syndrome
characterized by a significant decrease in red blood cells and physical abnormalities,
including craniofacial malformations. DBA is known to result from heterozygous mutations
in ribosomal protein (RP) genes, with over 20 causative genetic mutations identified to date.
However, approximately 40% of patients do not exhibit these mutations, suggesting thepresence of other causative genes or mechanisms. Furthermore, many aspects of DBA,
including treatment, genetic diagnosis, and the mechanisms of onset, remain unclear. We
have previously developed a comprehensive protein identification and quantification
method by Mass spectrometry. We conducted large-scale trans-omics analysis of humaninduced pluripotent stem cells (hiPSCs) and somatic cells. While mRNA remained constant
between these two cell-types, the analysis revealed 236 genes with significantly higher
protein in hiPSCs including nearly all reported DBA genes. We generated hiPSCs derived fromthe patients with high-frequency mutations in DBA genes such as RPS19, RPL11, and RPL5.
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These DBA-hiPSCs showed defective differentiation potential to early mesoderm.
Furthermore, while the protein levels of RPs in DBA-hiPSCs were comparable to those fromhealthy hiPSCs, they decreased during mesodermal differentiation. These findings suggestthat the DBA genes undergo different quantity control between hiPSCs and differentiatedcells. Importantly, this regulation occurs not during transcription but post-transcriptionally.
The data also imply abnormalities in the regulation of protein amount (proteostasis) during
differentiation and development in DBA patients. This study could propose a novel
mechanism to the onset of DBA through the regulation of proteostasis, and lead to a better
understanding of common principles underlying mutation-independent DBA pathogenesis.
Yin, Jiang-An

University of Zurich, CH

QUADRUPLE-GUIDE HUMAN GENOME-WIDE ARRAYED CRISPR LIBRARIES AND IPSC-BASEDRESEARCHES
Unbiased genetic screening is a foundational technique greatly advanced by CRISPR
technology. While current CRISPR screens utilize pooled guide RNA libraries with limited
applicability to selectable phenotypes, arrayed CRISPR libraries significantly expand thescope of CRISPR screens. However, generating arrayed CRISPR libraries is challenging andtheir availability is limited. Utilizing a newly developed massively parallel plasmid cloning
methodology, we have constructed genome-wide arrayed libraries for human gene ablation(19,936 plasmids), activation, and epigenetic silencing (22,442 plasmids). Each plasmid
encodes an array of four non-overlapping sgRNAs designed to accommodate most human
DNA polymorphisms. We achieve perturbation efficacies of 75–99%, 76–92%, and ~10,000fold in deletion, silencing, and activation experiments, respectively. Our studies havedemonstrated that our new libraries surpass existing resources in both superiority andversatility across various systems, particularly induced pluripotent stem cells (iPSCs) andorganoids. Given that our libraries unlock vast biological frontiers previously inaccessible andthe widespread utility of iPSC technology, we anticipate that the synergistic integration ofthese tools will significantly advance both fundamental and translational research inbiomedicine.

19Session 4: Translational Applications
Nakamura, Emiri

Kyoto University, CH

TURBULENCE ENHANCES MITOCHONDRIA DELIVERY IN MEGAKARYOCYTE MATURATION,LEADING TO BIOGENESIS OF HIGH QUALITY OF IPSC-PLATELETS
Platelet (PLT) products, limited by blood donations, have a short shelf life of 4-5 days due toits efficacy limitation and risk of bacterial growth at room temperature storage. To addressthese issues, we proposed two breakthrough technologies to supply artificial PLTs: 1.) selfrenewing megakaryocyte progenitors established from human induced pluripotent stem
cells (iPSCs) as a master cell bank, which we named immortalized megakaryocyte progenitor
cell lines (imMKCLs) (Nakamura, Cell Stem Cell, 2014). 2. turbulent-flow bioreactors thatenabled high yield production of competent iPSC-PLTs from imMKCLs (Ito, Cell, 2018). We
herein found turbulence increases CD42b (GP1bα) expression, a marker for PLT function, in
iPSC-PLTs, which inversely correlated with phosphatidylserine exposure levels andsubsequent CD42b shedding. We identified two populations of iPSC-PLTs: CD42b positive (+)
and negative (-), where mitochondria activity was detected only in CD42b+ iPSC-PLTs. We
also demonstrated using GPIX knockout imMKCLs or applying a mitochondrial oxidative
phosphorylation uncoupler, that mitochondria activity is an upstream regulator of
extracellular CD42b expression on iPSC-PLTs. Moreover, inhibiting mitochondrial fission also
decreased CD42b+ iPSC-PLTs, strongly suggesting that CD42b- iPSC-PLTs result from
impaired mitochondria delivery. Interestingly, we observed turbulence exposure only during
the latter 4 days of the total 6 days of maturation is crucial for mitochondria delivery.
Meanwhile, turbulence diminished F-actin levels by 20-30%, as detected via flow cytometry,
whereas complete inhibition of F-actin lowered mitochondria inclusion in resulting iPSCPLTs, implying that lower F-actin within an optimal range may be involved in mitochondriadelivery in imMKCLs to iPSC-PLTs. From these findings we propose that turbulence induces
the fine-tuning of mitochondria distribution via an actin mediated mechanism, to yield highquality iPSC-PLTs.
Clément, Blandine

ETH Zurich, CH

NERVE MODEL TO STUDY THE DIVERSE ELECTROPHYSIOLOGICAL PROPERTIES OF HUMAN
IPSC-DERIVED SENSORY NEURON SUBTYPES
Neuropathic pain results from various types of nerve damage and is often inadequately
treated. Most available in vitro tools to search for new therapeutic approaches fail togenerate human translatable results. Nociceptors are a specialized subpopulation of sensory
neurons conveying the perception of pain and are morphologically and functionally distinct
from other neurons. The individual sensation of pain can be triggered by different
subpopulations and may vary between patients. However, currently available painkillers
against neuropathic pain have no cell specificity resulting in unwanted side effects. Thus, thedevelopment of tools that can differentiate nociceptive fibres would enable a more targetedcompound screening. In this work, we present a multi-compartment nerve model platform
combining a CMOS-based high-density microelectrode array with a polydimethylsiloxane(PDMS) guiding microstructure that aims to capture the electrophysiological responses of20

individual dorsal root ganglion (DRG) subtypes. Human iPSC-derived sensory neurons were
cultured at low density inside the microstructure and only a few axons grew in multiple
parallel 4 x 10 μm microchannels before converging to a bigger bundle-forming channel. Thisconfiguration allowed the measurement of propagation speeds and stimulation-induced
responses of close-to-individual axons in microchannels after stimulating electrically thefiber bundle. We observed different electrophysiological profiles highlighting the diversity ofnociceptive fibres within a single network, suggesting that such a platform might be suitableto evaluate nociceptor-specific drug response for a given subtype of sensory neurons.
Jones, Lewis

ETH Zurich, CH

USING METAMATERIALS AND CARDIAC TISSUE ENGINEERING TO ENGINEER ROBUST AND
CONTRACTILE CARDIAC TISSUE PATCHES
Ventricular Septal Rupture (VSR) is a challenge in cardiac medicine with a high mortality rateof 45-60%. Current treatment methods use bovine pericardial patches (BPPs), which are noncontractile, tend to calcify over time, and fail to integrate effectively with the myocardium.
Therefore, patients do not tend to fully recover cardiac function. To address theselimitations, we are engineering a cardiac tissue patch that uses human stem cell-derived
cardiomyocytes in hydrogel, reinforced with a metamaterial lattice. This approach allows us
to tune the patch mechanical properties and contractility, while enabling stable implantation
within the intraventricular space. Here, we will showcase our current results onmetamaterial design and manufacturing, mechanical characterization (tunable stiffness),and biological characterization (biocompatibility, cell maturation, and tissue contractility). In
summary, we will show how metamaterials can be combined with engineered cardiac tissues
to fabricate centimeter scale, three dimensional, and implantable cardiac tissues.

21Core Facilities
Abidi, Affef

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THE IPSCORE ZURICH
The iPSC Core Facility (iPSCore) is a technology platform of the University of Zurichestablished to provide a wide range of services from induced pluripotent stem cell (iPSCs)
including reprogramming to differentiation, hands-on training, and project planning to thelife science community in Zurich and beyond. Currently, following protocols have been
established and are available for iPSC-derived cells: cardiomyocytes, smooth muscle cells,
endothelial cells neural progenitor cells, multiple types of neurons, astrocytes andmacrophages. Additional differentiation protocols are being established upon demand, suchas sensory, motor neurons and brain organoids.
We have recently completed our portfolio, and extend the possibilities of disease modellingusing iPSC, by offering CRISPR editing as a service, we can generate knock-out lines, mutation
correction/insertion in control/disease lines and transgenic/reporter lines. The mission of
the iPSCore is to empower its users with access to the state-of-the-art technologies in thefields of gene editing and stem cells, thus, enable the creation of research models best fit for
the unique need of each lab. We provide the full range of service, from the idea to the final
product, including the highest standard of quality control.
Jirkov, PaulinMore info here

UZH 3R AWARD 2024: CALL FOR NOMINATIONS IS OPEN!
The UZH 3R Award is intended to highlight the efforts and successes of UZH staff in the fieldof 3Rs (replacement, reduction, refinement of animal use in science), both within andoutside UZH, and thus promote the 3R principles at UZH.
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The prize is awarded to a member of the university (individual or team) who makes
a significant contribution in the field of the 3Rs.
Candidates must be employed or enrolled at UZH.
Candidates can be nominated by any UZH member.
The contribution may take the form of proposals to modify existing researchtechniques or innovative approaches that improve animal welfare in animal
experimentation or reduce or replace the use of animals.
Published scientific papers or publicly available preprints by UZH researchers as wellas education and training projects and initiatives for the implementation of 3R
methods by UZH members at UZH can be nominated. The publication, project or
initiative should have been published or taken place in the year prior to the year of
the award.
The award is endowed with 5000 CHF and is presented annually.
Deadline for nomination is 15.06.24, 23:59.

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Othman, Alaa

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FUNCTIONAL GENOMICS CENTER ZURICH (FGCZ): 20 YEARS AT THE FRONTIER OF MULTIOMICS SUPPORT FOR BASIC AND APPLIED BIOMEDICAL RESEARCH
The Functional Genomics Center Zurich (FGCZ) is a research and training platform of the ETH
Zurich and the University of Zurich, offering the latest technologies and key expertise formulti-omics research.
The FGCZ is leveraging Next Generation Sequencing (NGS) technology to support research
projects from population-scale profiling to measuring individual molecules in single cells.
Bringing together the latest sequencing technologies, automation solutions, and expertise inhandling ultra-low input material allows the scalable profiling of mutations, epigenetic
states, transcriptomic responses, etc., in virtually any tissue. We further exploit our powerful
NGS platforms and know-how by branching out into the field of genome editing andsupporting CRISPR screening and Off-Target analysis for next-generation genome editing
tools.
Penton Ribas, David

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THE ELECTROPHYSIOLOGY CORE FACILITY (E-PHAC) AT UZHThe Electrophysiology Facility (e-phac) offers expertise, instrumentation, and support in thearea of in-vitro electrophysiology to researchers from UZH and associated institutions. We
combine in-vitro electrophysiology with the advantages of image-based techniques to study
bioelectrical signaling in cells and tissues. We utilize manual and automated patch clamp, as
well as High-Density Microelectrode Arrays and other electrophysiological techniques, toassess the bioelectrical properties of ion channels, transporters, cells, and tissues. We
provide training for researchers in the use of these techniques and make a user Lab available,
or offer these techniques as a service. Please, get in touch with us to set up an experimental
plan for your project.
Stefanic, Sasa

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NANOBODIES, HIGHLY SPECIFIC ANTIGEN-BINDING DOMAINS AS AN ALTERNATIVE TO
CLASSICAL ANTIBODIES FOR USE IN RESEARCH, DIAGNOSTICS, AND THERAPY
Nanobodies are recombinantly produced antigen binding domains derived from antibodies
that that lack light chains, termed heavy chain-only antibodies, which naturally occur in all
camelid species and some cartilaginous fish. The main advantages over conventional
antibodies is in much smaller size and solubility that reflects in superior tissue penetration,
excellent thermal stability, and ease of expression and selection in vitro, whilst maintaining
the same binding affinities as conventional antibodies. In addition, nanobodies are often able
to recognise epitopes which are not accessible to conventional antibodies. Nanobodies canbe easily genetically modified and recombinantly produced in unlimited amounts using any
expression system (bacteria, yeast, insect cells) and further characterized in analytical andfunctional assay of choice. The Nanobody Service Facility of the University of Zurich has
specialised in the production of nanobodies by immunization of alpacas with the target
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protein, followed by the generation of a phage display library and in vitro selection of targetspecific binders. Through our service we make the nanobody technology available to the Life
Science Community (https://www.nsf.uzh.ch/en.html).
Applications in research range from immunohistochemistry, live imaging, pull-down assays,
functional analysis of protein-protein interactions, modulation of protein functions in thecell, or as mediators of specific protein knock-out. To date many nanobodies were identified
with potential to inhibit tumour growth and shown a great potential in non-invasive cancer
imaging or immunotherapy, but also against viral and bacterial infections, and neutralizing
toxins. The first nanobody was EMA and FDA approved for human therapy, and many moreare currently in the late phase of clinical trials. Highly diverse nanobody libraries also offer a
powerful high-throughput tool for development of diagnostic tests.

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Poster Abstracts
1 . Achón Buil, Beatriz

University of Zurich, CH

CRISPR ACTIVATION SCREEN IN HUMAN IPSC-DERIVED NEURAL PROGENITOR CELLS TO
ELUCIDATE MOLECULES ENHANCING THEIR MIGRATION TOWARD STROKE TISSUE
Stem cell-based therapy has emerged as a promising strategy for treating stroke, but it stillpresents several challenges, such as the route of administration. In a clinical setting,endovascular delivery of cells is preferred to intracerebral engraftment. However, circulatingstem cells must cross brain barriers and might get trapped in peripheric organs, resulting in
an insufficient number of cells in the ischemic lesion. To optimize stem cell migration acrossbrain barriers, we set out to mimic immune cells as they extravasate into the brain
parenchyma following stroke. For instance, since the chemokine CXCL12 is released after
stroke, we transduced human xeno-free iPSC-derived neural progenitor cells (NPCs) tooverexpress its receptor (CXCR4) which can be found on the surface of leukocytes.
Transduced NPCs migrated more efficiently than wild-type NPCs towards 10nM CXCL12 in a
Boyden chamber assay. Although this is a promising result, it is a biased approach thatneglects the complexity of the brain barrier extravasation. Therefore, we will develop a
CRISPR activation (CRISPRa) screen to determine the optimal targets for crossing brain
barriers after stroke. NPCs containing the CRISPRa system will be first tested in vitro byadding brain lysates to the Boyden chamber assay, or by performing adhesion assays. Top
candidates will be then injected endovascularly in mice following stroke. NPCs that arrive at
the ischemic lesion will be further analyzed to elucidate which molecules enhance theirmigration across brain barriers. The generation and preclinical validation of engineered NPCs
with improved brain homing capacity will improve the efficacy of stem cell-based therapies
for treating stroke via endovascular injection.
2 . Amimoto, Naoya

Kyoto University, JP

RECONSTRUCTION OF CORTICOSPINAL TRACTS BY TRANSPLANTATION OF HIPS CELLDERIVED CEREBRAL ORGANOIDS
Cerebrovascular disease is the second most common cause of long-term care needs. In
addition, the annual medical cost of cerebrovascular diseases is 1.8 trillion yen, and thedevelopment of treatment methods is urgently needed from a medical and economic
perspective. Cerebral infarction accounts for 80-85% of all strokes, which cause motor
dysfunction due to damage to the corticospinal tracts (CST) that connect the cerebral motor
cortex and the spinal cord. Various treatments for cerebral infarction, such as drug therapy
and rehabilitation, are available, but the fact that many patients remain suffering fromaftereffects makes it clear that the current therapies are insufficient. Therefore, as a novel
therapeutic strategy, there are high expectations for reconstruction of corticospinal tracts
by transplantation of pluripotent stem cell-derived neural cells such as iPS cells. The CST is
responsible for the transmission of motor information, including fine motor skills, one of themost important motor movements in humans, but there are no reports demonstrating thereconstruction of the CST. Therefore, our laboratory aims to reconstruct CST by celltransplantation. We have already confirmed that transplantation of human iPS cell-derived
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cortical neurons into the cerebral cortex of mice results in the elongation of neuro-axons
along the CST to the spinal cord and improvement of motor function1,2). However, previous
studies have not confirmed "graft-host connections" and "direct contribution to motor
function." Therefore, in this study, I would like to demonstrate "graft-host connections" and"direct contribution to motor function" by cell transplantation using the rabies virus andDesigner Receptor Exclusively Activated by Designer Drugs (DREADD). In fact, the study of
input from the host to the graft revealed that it received input from the same locations as
the actual input source, such as the cerebral cortex and thalamus. I believe that thesetechniques will demonstrate the integration of transplanted cells with the host's neuralnetwork of the brain and prove "reconstruction of the corticospinal tract by celltransplantation.
3 . Amos, Giulia

ETH Zurich, CH

STUDYING LOCAL PLASTICITY IN LOW-DENSITY IPSC-DERIVED NEURONAL NETWORKS
To this day, we lack a basic understanding of how the human brain retrieves, processes andstores information. One particular challenge is unraveling how local plasticity mechanisms
shape information transfer between neurons. In literature, we can find various plasticitybased learning paradigms, such as the Spike-Timing Dependent Plasticity or the BienenstockCooper-Muncro synaptic learning rule. However, many of these paradigms faceexperimental and functional limitations as they exclusively encode spike timing or firing rate,and it is still unclear how they relate to each other. This might be attributed to the lack ofwell-defined model networks of neurons that can be reproducibly and quantitatively studied
over an extended period of time. We introduce a bottom-up platform that combines
engineered in vitro neuronal network models with microelectrode arrays (MEAs) to
overcome this challenge. In this platform, human induced pluripotent stem cell (hiPSC)derived neurons are cultured within custom-designed polydimethylsiloxane (PDMS)
microstructures placed on top of the MEAs to engineer neuronal networks with constrained
connectivity and compartmentalisation. These topologically constrained networks display
complex spatiotemporal dynamics, making them valuable experimental tools to represent
neuronal in vivo networks. Consequently, the platform enables to study plasticity rules inisolated neuronal networks with reduced complexity, while providing high reproducibility
and throughput. We show that we can modulate the network activity using stimulation
protocols that could potentially evoke spike-time dependent plasticity effects. Looking
forward, we aim to test the learning paradigms further by simplifying the networks tocontain 1-3 neurons. These investigations may provide insight into the levels of complexity
required for learning within an in vitro laboratory setting and could provide a valuable
complementary approach to in vivo methods.
4 . Budny, Vanessa

University of Zurich, CH

THE ROLE OF APOLIPOPROTEIN E (APOE) ISOFORMS IN NEURAL CELL PHYSIOLOGY
Alzheimer’s Disease (AD), characterized by the accumulation of beta-amyloid plaques andneurofibrillary tangles, is the most common age-related neurodegenerative disorder. The
APOE4 allele is the major genetic risk factor for AD while APOE3 is defined as average risk
and APOE2 is protective. Despite recent advances, the fundamental role of different APOE
26

alleles in brain homeostasis is still poorly understood. Here, we aim to uncover the functionalrole of APOE2, E3 and E4 in human neural cells and how this relates to AD pathophysiology.
We differentiated APOE-isogenic iPSCs (APOE4, E3, E2 and APOE-knockout (KO)) tofunctional astrocytes (“iAstrocytes”) and neurons (“iN cells”). iAstrocytes were analyzed forproteomic profiles using unlabeled mass spectrometry. GO enrichment analysis showed
genotype differences in various cellular pathways including inflammatory signaling andenergy metabolism. Accordingly, APOE4 iAstrocytes displayed the highest release ofcytokines, while APOE2 and APOE-KO iAstrocytes secreted the lowest amounts.
Furthermore, APOE4 iAstrocytes showed lower capacity for uptake of beta-amyloid andglutamate, while uptake was highest in APOE2. To investigate the role of APOE in energy
metabolism of both, iAstrocytes and iN cells, we used the seahorse assay. Interestingly, total
as well as mitochondrial ATP production were highest in APOE4 iAstrocytes and iN cells
compared to the other genotypes, while no difference was observed in iPSCs suggesting acell-type-dependent effect. Novel, single cell-based functional assays will be applied tocharacterize the APOE-dependent regulation of neural energy metabolism in more detail.
We show that APOE plays a major role in several physiological and metabolic processes inhuman neural cells with APOE4-expressing iAstrocytes being pro-inflammatory and deficientin beta-amyloid and APOE4 iN cells and iAstrocytes being affected in their bioenergetic state.
5 . Carestiato, Silvia

University of Turin, IT

HUMAN IPSCS-DERIVED NEURONAL MODEL TO STUDY THE PATHOGENIC MECHANISMS
BEHIND TANGO2-DISEASEThe TANGO2 gene encodes for the homonymous protein mainly involved in vesiculartrafficking, mitochondrial oxidation, and cellular homeostasis. While its subcellular
localization is unclear, TANGO2-biallelic LoF variants are associated with an autosomal
recessive disorder characterized by cardiac arrhythmia, metabolic crises, rhabdomyolysis,
and kidney damage. A neurological component is also observed, including progressive
neurodegeneration, developmental delay, and cerebral atrophy. Although a possible role inneuronal migration has been suggested, the effect of TANGO2-loss in neurons has neverbeen investigated, constituting a major gap in understanding the disease.
Human induced Pluripotent Stem Cells (hiPSCs) were generated from two clinicallydiscordant TANGO2-affected siblings to derive neural rosettes (NRs), neural progenitor cells(NPCs) and mature neurons. Analysis of NRs showed a significative area reduction (~45%)
and an impaired morphological organization in the severely affected patient. Furthermore,in the same patient, the wound-healing assay demonstrated altered migration in NPCs,
supporting the previously reported role of TANGO2 in migration. Additionally, during thematuration of cortical neurons, an increase in cell death was also observed. In this regard,
the Reactive Oxygen Species (ROS) assay showed an oxidative stress condition supporting
the involvement of TANGO2 in mitochondrial functions and cellular homeostasis.
In contrast, no significant differences were found in NRs, NPCs, and mature neurons in theasymptomatic patient compared to controls. In conclusion, we demonstrated the role of
TANGO2 in early-stages of neurodevelopment, particularly in neural tube formation, cell
migration, and neuronal maturation. Considering the peculiar siblings' case, this study
suggests potential protective gene/genetic modifiers influencing patient phenotypes,
highlighting the need for further transcriptomic and proteomic analyses for future therapies.
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6 . Carreira, OlgaNOVA Medical School Research, PORTMODELLING RETINAL DEGENERATIVE MECHANISMS USING 3D RETINAL ORGANOIDS
The major causes of human vision loss and blindness are due to degenerative retinal diseases
that involve function loss or death of specific retinal cell types. Recent studies have foundretinal abnormalities in Parkinson’s disease (PD) patients and retinal deposition of α
synuclein (α syn). The development of reliable models to study the human retina has
improved and 3D models like organoids derived from hiPSCs have been described torecapitulate the cellular organization and light sensitivity of the human retina. Here, we aim
to study the effects of α syn overexpression in retinal degeneration, by generating retinalorganoids derived from a PD-patient iPSC line with a triplication of the α syn gene (SNCA)
and from a control cell line. Preliminary results indicate that PD-patient retinal organoids are
efficiently generated and self-organized compared to control organoids. At different
differentiation stages (days 80 and 120) PD-patient organoids revealed higher α syn protein
levels and its phosphorylated form at serine 129. In parallel, we observed increased protein
levels of antioxidant enzymes, such as catalase and superoxide dismutases (SODs) 1 and 2,
together with increased mRNA levels of pro-inflammatory cytokines like interleucine 6 (IL6).
Additionally, members of the B cell lymphoma 2 (BCL2) family like BCL2, Apoptosis Regulator(BAX) and BCL2 antagonist/killer 1 (BAK1) also displayed higher mRNA levels in PD-patientorganoids, as well as higher levels of cleaved Caspase-3 (c-Casp3), indicating disfunction
related with the mitochondrial pathway of apoptosis. Overall, this work supports 3D retinalorganoids as a reliable model to study the molecular mechanisms involved in retinaldegenerative diseases and further validate them as a tool for developing targeted andinnovative therapeutic strategies against oxidative stress and degeneration.
7 . Collo, Linda

University of Genova, IT

CHEMICAL STIMULATION OF NEURAL NETWORKS DERIVED FROM HUMAN INDUCEDPLURIPOTENT STEM CELLS
In vitro models combined with human induced pluripotent stem cells (hiPSCs) are a widelyused tool to investigate the complex mechanism of the human brain. Indeed, the use of
hiPSCs-derived neuronal networks allow to create in vitro patient-specific models that mostclosely resembles the processes of the human system, providing a reliable model to test
neuroactive properties of different compounds. This study investigates how different drugs
affected the electrophysiological activity of in vitro systems composed by excitatory (E) andinhibitory (I) neurons derived from hiPSCs. Our experimental model included three different
culture configurations: 100% excitatory neurons (100E), 75% excitatory and 25% inhibitory
neurons (75E25I), and 100% inhibitory neurons (100I). To these neuronal networks, 2-amino5-phosphonovaleric (APV), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and bicuculline(BIC) were administrated. The recordings of the spontaneous and chemically stimulated
electrophysiological activity were performed by exploiting Micro-Electrodes Arrays (MEAs).
The administration of APV and CNQX resulted in an unaffected firing activity, while led to adecrease of the bursting and network bursting activity of both 100E and 75E25I
configurations. On the other hand, BIC led to a significant increase of the firing activity of the
100I and 75E25I, and of the bursting and network bursting activity of the 75E25I
configuration. This comprehensive investigation sheds light on the different effects of APV,
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CNQX and BIC on the electrophysiological dynamics of neuronal networks with different
excitatory-inhibitory compositions. These findings contribute to our understanding of druginduced modulation in neuronal circuits and provide valuable insights for potential
therapeutic investigations in neurological disorders.
8 . Dodi, Lorenzo Davide

ETH Zurich, CH

INVESTIGATING ADHD BIOMARKERS USING HIPSC DERIVED NEURONS ON MEAS
Attention-deficit hyperactivity disorder (ADHD) is among the most prevalentneurodevelopmental disorders worldwide, with up to 7% of children and 4% of adults being
affected. Psychostimulants are currently the first-line treatment in childhood and adult
ADHD. Despite their high efficacy, approximately 30% of children and over 50% of adults do
not respond sufficiently to psychostimulants, and a substantial proportion of patients stoptreatment due to side effects. Predictive markers for an early identification of potential nonresponders currently do not exist, leading to unnecessary drug exposure for many patients.
Current state-of-the-art approaches to study ADHD have significant limitations: animalmodels fall short in representing complex human mental disorders, and neuroimaging does
not offer access to single-cell data at high temporal resolution, which is a key prerequisite
for in-depth network characterization. The main goal of this project is to perform
electrophysiological characterization of disease-induced effects in patient-derived cells athigh spatiotemporal resolution, combining human stem cell biology with cutting-edge highdensity microelectrode array (HD-MEA) technology. Human iPSC-derived neurons retain theunique genetic signatures of their donors and enable the study of complex polygenic
disorders, such as ADHD. HD-MEA technology provides access to network activity at highspatiotemporal resolution, ranging from subcellular compartments through individual
neurons to entire networks. This combination potentially enables data-driven phenotypic
screening to facilitate medication assessment and support the development of personalized
treatment strategies. It may contribute to improving treatment outcomes of individual ADHD
patients, reducing individual suffering, and developing novel treatment options.
9 . Duré, Clara

University of Zurich, CH

IN VIVO SINGLE-CELL RIBOSOME PROFILING TO INVESTIGATE TRANSLATIONAL REGULATIONIN AGED EPIDERMAL STEM CELLS
Adult stem cells are characterised by a tightly regulated and globally suppressed translation.
With advanced age, stem cell function declines and the capability to self-renew andregenerate a tissue decreases. There is direct evidence that translation plays a major role inageing since low levels of protein synthesis are associated with longevity. However, how thetranslational landscape of stem cells is altered during ageing remains largely unknown. Here,we employ in vivo single-cell ribosome profiling of the mouse epidermis to unravel the roleof translational regulation in epidermal stem cells and the consequences of translational
deregulation on skin regeneration. By optimizing the recently published single-cell ribosome
profiling protocol specifically for in vivo epidermal stem cells, we comprehensively explore
the translational landscape of young and aged epidermis at single-cell resolution, elucidatingcell-type-specific translational programs during ageing. Furthermore, by coupling single-cellribosome profiling with single-cell RNA sequencing, we uncover strongly deregulated29translation specifically in aged epidermal stem cells, which contrasts with the prevalenttranscriptomic changes observed in aged differentiated epidermal cells. Additionally, aged
epidermal stem cells show elevated expression of inflammatory response genes, that aretranslationally regulated. Systemic inflammation is a hallmark of ageing and together with
our findings, it could be under translational control in ageing stem cells. Together, ourfindings advance our understanding of how deregulated translation contributes to agerelated systemic inflammation, stem cell exhaustion and might pave the way for improving
stem cell fitness, which is particularly relevant for wound healing of aged skin.
10 . Elsafadi, SaraETH Zurich, CH

GENERATION OF INDUCED PLURIPOTENT STEM CELLS (iPSCS) FROM ROE DEER (CAPREOLUS
CAPREOLUS) EMBRYONIC FIBROBLASTS VIA EPISOMAL VECTORS
Embryonic diapause, a period of developmental delay, remains an enigmatic process despite
being widely observed among various mammalian species. This phenomenon was first
documented in the European roe deer (Capreolus capreolus) in 1854. It features a distinctive
extended phase lasting about four months, by which slow but continuous embryonic growthoccurs. The collection and in vitro production of roe deer embryos for research purposes are
fraught with field work difficulties, positioning induced pluripotent stem cells (iPSCs) as apromising research tool. The iPSC provides an innovative means to delve into thecomplexities of cellular potency and embryonic mechanisms of developmental delay. In thisstudy, we strived to generate iPSC lines from roe deer embryonic fibroblasts utilizing
episomal vectors encoding crucial pluripotency factors - Oct3/4, SOX2, KLF4, L-MYC, and
LIN28 - in a feeder- and serum-free milieu. Although our initial analyses reveal that OCT4
expression in the reprogrammed cells likely originates from the introduced vectors rather
than endogenous activity, early results are encouraging, indicating the potential for
reprogramming roe deer fetal fibroblasts with human factors under these conditions.
Comprehensive cell characterization is pending to ascertain their true stem cell capabilities
and reliance on these vectors. Developing an iPSCs model for the roe deer promises tounravel the cellular and molecular intricacies of embryonic diapause. This advancement has
the potential to significantly advance our comprehension of mammalian reproductive anddevelopmental mechanisms, enriching the field of stem cell research by providing insight
into species-specific pluripotency and cell cycle regulation.
11 . Eschment, Melanie

University of Zurich, CH

AN IPSC-DERIVED CEREBRAL ORGANOID MODEL FOR CEP290-ASSOCIATED
NEURODEVELOPMENTAL CILIOPATHIES IDENTIFIES ANOMALIES IN CILIARY MORPHOLOGY
Ciliopathies are a group of human Mendelian disorders caused by dysfunction of primarycilia, small ubiquitous sensory organelles protruding from the surface of most cells, required
for signal transduction. These disorders are associated with central nervous system (CNS)
anomalies, particularly exemplified by the neurodevelopmental disease Joubert syndrome(JBTS). JBTS is characterized by a highly specific mid-hindbrain malformation whose
underlying pathomechanism remains unclear. Moreover, the presence of non-structural CNS
defects such as seizures or intellectual disability implies a role for cilia in neuronal function
beyond transmission of developmental signaling pathways. To understand the role of cilia
30and of JBTS genes in the CNS, we are generating iPSC-derived in vitro models for JBTS. We
established a biobank for JBTS through generation of CRISPR-engineered isogenic humaninduced pluripotent stem cell (hiPSC) lines that carry mutations in selected JBTS-associatedcilia- related genes such as the ciliary gene CEP290. To elucidate the consequences ofmutations in CEP290 on brain development, we differentiated control, CRISPR-edited andpatient-derived hiPSCs into 3D cerebral organoids. We find that CEP290-mutated hiPSCs candifferentiate into cerebral organoids and single-cell RNA sequencing confirmed that theoverall generation of a variety of neuronal cell populations does not appear to besignificantly impacted up to day 80. We do, however, observe an increased propensity togenerate choroid plexus in mutant organoids compared to controls and a disorganization ofcortical plate units. Furthermore, we observe striking morphological anomalies of a subset
of primary cilia in the lumen of cortical plate units generated, which is a consistent finding
throughout our genetically engineered and patient-derived cerebral organoids. Altogether,
this study represents an advancement in understanding JBTS in the context of human brain
development.
12 . Fujisawa, Miwako

University of Bern, CH

INTRINSIC BLOOD-BRAIN BARRIER DYSFUNCTION CONTRIBUTING TO MULTIPLE SCLEROSISPATHOGENESIS
Blood-brain barrier (BBB) breakdown is amongst the earliest pathological hallmarks
observed in multiple sclerosis (MS). The mechanisms leading to BBB dysfunction are
incompletely understood and are generally thought to be a consequence of the autoimmune
neuroinflammatory process in MS. Investigating BBB dysfunction in MS is hampered by thelimitation of MS tissue samples. Therefore, we made use of human induced pluripotent stem
cells (hiPSCs) from MS patients and healthy controls (HC) and established in vitro models of
the BBB by differentiating hiPSCs into brain microvascular endothelial cell (BMEC)-like cells
using the extended endothelial cell culture method (EECM) recently developed by us. EECMBMEC-like cells from MS patients showed impaired barrier properties and an inflammatory
phenotype when compared to those derived from HCs. Comparing the transcriptional profileof the first EECM-BMEC lines we found 438 significantly differentially expressed genes in
non-stimulated MS- versus HC-derived EECM-BMEC-like cells and 343 differentiallyexpressed genes in cytokine-stimulated MS- versus HC-derived EECM-BMEC-like cells. Thesefindings underscore that intrinsic dysfunction of the BBB contributes to MS. To further
substantiate these findings and to explore the underlying mechanisms we have now
established hiPSCs from 3 additional HC and 8 additional MS patients and differentiatedthem into EECM-BMEC-like cells. EECM-BMEC-like cells from two additional MS patientsprior to diagnosis (radiologically isolated syndrome: RIS) and at the time of diagnosis withMS are included to determine if BBB dysfunction establishes prior to clinical signs of MS.
Characterization of these data will be presented. The differences in transcriptional profileand phenotype of these MS- versus HC-derived EECM-BMEC like cells will be presented.
13 . Gatta, Beatrice

University of Zurich, CH

INVESTIGATING THE ROLE OF DEFINED TDP-43 STATES IN HEALTHY AGING AND ALS PATIENTDERIVED NEURAL NETWORKS
31TAR DNA-binding protein 43 (TDP-43) is the main aggregating protein in several
neurodegenerative diseases including Amyotrophic Lateral Sclerosis (ALS) and
Frontotemporal dementia (FTD). Yet the mechanisms that trigger the transition of TDP-43
from a highly regulated protein to a non-functional and pathological form that in turn causes
toxicity and neuronal death remain unknown. An interesting hypothesis is that TDP-43 canexist in different functional states that allow the protein to facilitate its many functions incells. Some of these states could be less functional then others and therefore may initiate
the shift from a physiological to a pathological state of TDP-43 in disease. In order to decipher
the functionality of distinct TDP-43 states and to elucidate their possible role in driving
pathology, we use aging human neural networks established in our lab. Notably, this humanneural model consists of self-organized and multi-layer glia and neural cultures withsynaptically connected and electrophysiologically active neurons, which matures into a longlived functional neural network over time. An important advantage of this system is its highreproducibility and significant longevity that makes it ideal to study neurodegenerative
diseases such as ALS. Taking advantage of engineered iPSCs harboring ALS-linked mutations
as well as ALS patient-derived iPSCs, we want first to generate aging neural cultures and thenperform state-of-the art single-cell transcriptomic analysis as well as molecular andbiochemical assays on these long-lived cultures to investigate the emergence of diseaserelevant TDP-43 states and their impact on human neural cultures over time. Most
importantly, we aim at evaluating the long-term impact of TDP-43 states in human neurons
so as to understand how relevant pathological features develop in the course of time withthe attempt of recapitulating the late-onset events associated to TDP-43-causingneurodegenerative diseases. This research will allow us to confirm the possible role of
relevant TDP-43 states in ALS disease and to shed light on new possible pathways of toxicity
underlying TDP-43 pathology.
14 . Haenseler, Walther

University of Zurich, CH

HUMAN IPSC DERIVED MODELS TO EXAMINE ALPHA-SYNUCLEIN PATHOLOGYA Hallmark of Parkinson’s Disease is α-synuclein (αS) accumulation in aggregates called Lewy
bodies. To examine the pathogenic role of αS, we have generated induced Pluripotent StemCells (iPSC) from early onset Parkinson's Disease patients with SNCA A53T and SNCA
Triplication mutations as well as gene edited isogenic sets. We characterized iPSC derived
macrophages and found significantly reduced phagocytosis capability in SNCA Triplication
macrophages, but not A53T macrophages. This phagocytosis defect can be phenocopied byadding αS to the cell culture medium of control macrophages. We characterized iPSC derived
dopaminergic neurons and found increased αS aggregation and mitochondrial defects.
In ongoing projects, we use limited proteolysis followed by mass spectrometry (LiP-MS) to
identify structural changes in the proteome. We used SNCA KO iPSC derived cortical neuron
lysates and spiked them with monomeric and fibrillar αS to identify αS structure specific αSinteraction partners with LiP-MS. Furthermore, we are now characterizing the differences inthe structural proteome between healthy controls and the SNCA triplication patient and theconsequences of spiking αS fibrils into live neurons derived from control, SNCA KO and SNCA
Triplication iPSC. With these human iPSC models we have confirmed finding from animalmodels and patient samples, but also got novel insight into αS pathology. They are an
unlimited source of live patient material normally not available for disease modeling.
32

15 . Helfenstein, Carmen

University of Zurich, CH

SAFETY SWITCH SYSTEMS IN HUMAN IPSC-DERIVED NEURAL PROGENITOR CELLS TO ENSURE
SAFE APPLICATION IN STROKE
Ischemic stroke is a leading cause of severe long-term disability and death due to the limited
regenerative capacity of the brain. Stem-cell-based therapy holds promise for stroke
treatment, yet the proliferation potential poses a risk of tumorigenesis. Safety switch
systems, based on suicide gene therapy from tumor research, offer a potential solution by
inducing cell death specifically in modified cells. They consist of a cell-suicide-inducing
transgene and a specific molecule that, when combined in the cell, cause cell death. Most
studied safety switch transgenes are herpes simplex virus thymidine kinase (HSV-TK) and
inducible Caspase9 (iC9) genes that induce cell death upon the administration of ganciclovir(GCV) or AP20187 respectively. Human iPSC-derived neural progenitor cells (NPCs) weretransduced separately with two lentiviral constructs containing HSV-TK or iC9, together with
firefly luciferase and a fluorophore reporter. We further plan to carry out a doubletransduction to remove all cells (iC9) or only dividing cells (HSV-TK). We determined in vitro
the optimal drug concentration to trigger cell death via bioluminescence andimmunostaining. In future experiments, modified NPCs will be intracerebrally injected into a
photothrombotic stroke mouse model followed by drug administration. NPCs will be tracked
in vivo via bioluminescence imaging, supplemented by post-mortem immunostaining forcomprehensive analysis. These findings represent a significant stride toward the safer
application of stem-cell-based therapy in treating stroke patients.
16 . Lloyd-Davies Sánchez, Daniel

University of Cambridge, UK

OF MICE AND MEN - CEREBRAL ORGANOIDS FOR THE STUDY OF BRAIN DEVELOPMENT
Cerebral organoids grown with minimally guided protocols are able to recapitulate aspectsof species-specific differences in timings of brain development which exist in-vivo.
Furthermore, these stem-cell derived models can be used to make inter-species comparisons
which would otherwise be technically or ethically inaccessible to experimentation. Being
able to properly understand human-specific aspects of brain development will require us tobe able to understand the phylogenetic context and how we and our developmental
trajectories differ from close relatives such as apes, and also more distantly related mammals
such as rodents. Indeed, there is much variation in development between species; and as alarge literature exists for mouse as an in-vivo model, understanding the implications of thisfor human brain development requires new models which can bridge this knowledge gap
across species. We adapted the cerebral organoid protocol, previously used on humanderived stem cells, for other species including mouse and observed intrinsically fasterdevelopmental trajectories and differentiation of neural fates compared to human cerebral
organoids. Additionally, prolonged culture at the air-liquid interface reveals establishmentof early neuronal layering and further cell-type differentiation, as well as neuron and axonal
connections and dynamics.
17 . Kasap, Pelin

University of Bern, CH

A HUMAN IPSC-DERIVED ISOGENIC MODEL OF THE NEUROVASCULAR UNIT TO EXPLOREBLOOD-BRAIN BARRIER DYSFUNCTION IN NEUROINFLAMMATION
33

The blood-brain barrier (BBB) is established by brain microvascular endothelial cells (BMECs)
that maintain their characteristics by continuous crosstalk with pericytes and astrocytes. BBB
breakdown and immune cell infiltration into the CNS are the earliest pathological hallmarksof multiple sclerosis(MS). The mechanisms of BBB breakdown in MS are largely unexplored
due to the lack of patient-derived BBB models. To address this, we employed human induced
pluripotent stem cell(hiPSC) technology to build an isogenic neurovascular unit (NVU) onchip that models the anatomical structure of the postcapillary venules, where the immune
cell infiltration into the CNS parenchyma takes place. We built the μSiM platform(microphysiological system enabled by a silicon nitride nanomembrane) that features
ultrathin, transparent, and permeable membranes forming a “blood compartment” linedwith hiPSC-derived BMECs and a “brain compartment” harboring hiPSC-derived pericytes
and astrocytes. The μSiM offers the simplicity of the TranswellTM system while enabling live
imaging, the use of different membranes with variable pore sizes, and the addition ofmodules converting it to a microfluidic or a tricellular culturing device. We produced hiPSCderived EECM-BMECs that are characterized by an endothelial transcriptome. EECM-BMECs
cultured in μSiM formed a uniform monolayer with mature tight junctions, developed barrier
properties, and showed cytokine-induced expression of adhesion molecules. We successfully
added hiPSC-derived pericytes and astrocytes to the μSiM, forming a fully isogenic NVU. We
demonstrated successful T-cell migration from the blood compartment to the brain
compartment in the μSiM under static conditions. Overall, our data demonstrate thesuitability of the μSiM device to study the function of the human NVU in health andneuroinflammation. The addition of a flow unit will finally allow the investigation of immune
cell trafficking across the NVU in an autologous manner.
18 . Kido, Jun

Kumamoto University Hospital, JP

PATIENT DERIVED IPS CELLS AS CELLULAR MODEL FOR BASIC RESEARCH INTO CITRINDEFICIENCY AND FOR DEVELOPMENT OF NOVEL THERAPIES
Citrin deficiency (CD) is an autosomal recessive disorder caused by a defect of the innermitochondrial membrane transporter citrin resulting from mutations in SLC25A13. A defectof citrin primarily affects the function of the malate-aspartate shuttle but has several
downstream metabolic effects impairing energy production due to hampered glycolysis,
gluconeogenesis, tricarboxylic acid cycle function and beta-oxidation, and impairs nitrogendetoxification in the urea cycle. The condition is prevalent in East Asia, but also present in
the Western world and hence considered a global disorder. Patients require as high-fat andlow-carbohydrate diet with supplementation of medium-chain triglycerides (MCT) although
this treatment has been introduced empirically without sound scientific evaluation.
Especially the mechanism of action of MCT in this condition has never been systematicallystudied. We made use of patient derived skin fibroblasts, which were reprogrammed into
iPS cells using standard protocols prior to differentiation into hepatocytes. These cells wereused for studies of energy metabolism yielding decreased ATP levels and increased
NADH/NAD+ ratios confirming their usefulness as CD model cells. For control, we used
cultured patient fibroblasts and the hepatoma cell line HepaRG. Finally, we evaluated theeffect of medium-chain fatty acids (MCFAs) on the ATP level and NADH/NAD+ ratios. Hereby,
we could show in preliminary experiments that MCFAs lead to an increase of ATP levels
indicating the improvement of energy production in disease iPSCs. In conclusion, this study
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shows the usefulness of patient derived iPSCs both as disease model for CD and as a tool forstudying novel treatments for this condition.
19 . Küchler, JoëlBIOLOGICAL NEURONAL NETWORKSFUNDAMENTAL LOGICAL OPERATIONSETH Zürich, CH
AS

COMPUTATIONALUNITS:

ACHIEVING

Artificial intelligence (AI) has proven to be effective in solving complex problems in fields
such as computer vision, natural language processing and medicine. However, in terms ofenergy efficiency, deep learning approaches are still outmatched by their biologicalcounterparts. Yet, the underlying fundamental mechanisms of information storage,
processing and computation of biological neuronal networks (BNNs) remain poorly
understood. In bottom-up neuroscience, we tackle this challenge by analyzing low-density,
engineered BNNs in vitro. Primary hippocampal rat neurons or human induced pluripotent
stem cell (hiPSC) derived ngn2 neurons are seeded on top of high-density microelectrodearrays (HD-MEAs), where their growth is confined by polydimethylsiloxane (PDMS)
microstructures. This approach allows us to record and stimulate BNNs to study the
propagation of information on a subcellular level. It further has been demonstrated that themean firing rate of a response induced by periodic continuous stimulation does not change
significantly over periods of multiple hours. This gives a basis for using BNNs as acomputational unit. In this work, we use PDMS microstructures with nanochannels which
can generate isolated feed-forward circuits consisting of two input and one output node.
Artificial inputs are encoded by stimulation patterns and the response of the middle node iscaptured with rate or temporal coding. By implementing different stimulation protocols, we
successfully achieve three fundamental logical operations: NOT, AND, and OR. Furthermore,these operations are robust to stochasticity in the input and reproducible across different
circuits. In addition, we compare stimulation-induced and spontaneous responses. Initial
results show similarities not only in terms of spike timing but also in the principal
components of the recorded signal shape. Looking ahead, this observation would allow
interconnecting small logic gates to do more complex operations.
20 . Maurer, Benedikt

ETH Zurich, CH

QUANTIFYING ENVIRONMENTAL EFFECTS ON SPIKING ACTIVITY IN ENGINEERED NEURALNETWORKSIn vitro networks of human induced pluripotent stem cell (hiPSC) derived neurons have thepotential to provide a more physiologically relevant, high-throughput platform to study
electrical and chemical modulation of brain function under healthy and diseased conditions.
However, variability between cultures due to their complexity and instable environmental
parameters can have detrimental effects on readout robustness and reproducibility. In ourbottom-up approach, small neural networks of reduced complexity are engineered byseeding hiPSC ngn2 neurons into polydimethylsiloxane (PDMS) microstructures on top of
microelectrode arrays (MEAs) to spatially confine the location of their soma and guideneurite growth. Repetitive stimulation of these networks yields stable and reproducible
spiking patterns in the evoked responses over days, which can be modulated by adding
drugs. We present a custom readout and incubation system, Inkube, with recording and35

closed-loop stimulation capability for 4 standard 60-electrode MEAs in parallel as well as fully
integrated temperature and CO2 control. The medium temperature is controlled individuallywith an immersed resistive temperature sensor. Additionally, a syringe pump based
perfusion system with a feedback sensor for the fluid volume in the wells is added. We
demonstrate the temperature dependence of spike transmission latency in patterned
networks of reduced complexity. It is crucial to isolate drug induced effects from variations
caused by fluctuating environmental parameters. With our approach we aim to increase
reproducibility in network studies and provide a robust drug screening tool that serves the
3Rs principle (replacement, reduction, refinement).
21 . Nunes, Carolina

ETH Zurich, CH

HUMAN ESC-DERIVED RETT SYNDROME ELECTROPHYSIOLOGY AND MOLECULAR
PHENOTYPIC SCREENING PLATFORM FOR THE DEVELOPMENT OF NEW THERAPIES
Rett syndrome (RETT) is a rare neurodevelopmental disorder characterized by an early
period of normal development, followed by a sudden loss of acquired psychomotor skills.
Most cases of RETT are caused by sporadic mutations in the Methyl-CpG-binding protein 2(MECP2) gene on the X chromosome, which is essential for normal development,
maturation, and function of neurons in the brain. Consequently, many new treatment
strategies for RETT try to target MECP2 expression levels and their downstream pathways –but as of today – there is no cure for RETT. For a deeper understanding of how mutations in
MECP2 relate to neuronal function and the observed range of symptoms in patients, we set
out to develop an integrated multimodal phenotypic screening approach. Here, we present
preliminary data obtained from 2 and 3D human ESC-derived control/ mutant neuronalcultures using a combination of functional and genomics assays, including high-throughput
high-density microelectrode array (HD-MEA) measurements, patch clamp recordings, andgene expression analyses. Preliminary data indicate that MeCP2-deficient neuronal cultures
show a higher division rate and lower levels of mature neuron markers (e.g., markers of
microtubule-associated protein 2 and 200 kD neurofilament), as well as higher neuronalfiring rates during early development. Generally, neuronal networks developed morerobustly in 3D-spheroid cultures. When fully functional, this platform will be adapted tohiPSC-derived cultures and used to probe phenotypic rescue approaches including
pharmacological and gene-therapeutic interventions aimed at restoring MECP2 expression
levels.
22 . Oryshchuk, Anastasiia

MaxWell Biosystems

LABEL-FREE FUNCTIONAL CHARACTERIZATION OF IPSC-DERIVED NEURONS AT SUBCELLULARRESOLUTION
In recent years, brain models derived from pluripotent stem cells have become afundamental tool for studying common neurological disorders, such as epilepsy, Alzheimer’s
disease, and Parkinson’s disease. The ability to measure the electrical activity of human iPSC
derived neurons in real time and label free can provide much needed insights into thecomplexity of the neuronal networks. Nowadays, combining single cell resolution with highthroughput physiological assays, which can potentially deepen our understanding of subtype
specific neuronal activity, is especially valuable and yet difficult to achieve. In this study, the36

MaxTwo System (MaxWell Biosystems, Switzerland), a multi well high density (HD) MEA
platform was used. MaxTwo HD MEA System allows in vitro extracellular recordings of action
potentials at different scales, ranging from network through single neuron to subcellularfeatures. Moreover, we showed the advantages of having an HD-MEA system featuring
26,400 electrodes per well, which are key to increase the statistical power of the datacollected from iPSC derived neurons over multiple days, weeks and to capture the smallest
neuronal signals. Finally, we present the Axon Tracking Assay, a tool for automated recording
and analysis of individual axonal arbors of many neurons in parallel. The Axon Tracking Assay
enables to measure action potential conduction velocity, axonal length, and number ofaxonal branches. With this unique method, we characterized the function and axonal
structure of different iPSC derived neuronal cell lines. Our HD MEA platforms and theextracted metrics, such as firing rate, spike amplitude, and network burst profile among
several others, provide an extremely powerful and user friendly approach for in vitro drug
screening and disease modelling.
23 . Parodi, Giulia

University of Genova, IT

INVESTINGATING THE EFFECT OF CHEMICAL AND ELECTRICAL MODULATION IN HUMANIPSCS-DERIVED NEURONAL NETWORKS COUPLED TO HIGH DENSITY ARRAYS
In the brain, the neuronal activity is kept and controlled by a precise and well-defined“Excitatory/Inhibitory balance” among neurons, which holds importance inneurodegenerative and neurodevelopmental disorders. With the final aim of generating a
reliable in vitro system of the human brain, in this work, we focused on the interplay
between excitatory glutamatergic (E) and inhibitory GABAergic (I) neurons in neuralnetworks derived from human induced pluripotent stem cells (hiPSCs), deepening the criticalrole of heterogeneity. We exploited high-density Micro-Electrode Arrays (MEAs)
characterized by 2304 electrodes to explore two neuronal culture configurations: 100%
excitatory (100E) and 75% excitatory / 25% inhibitory (75E25I) neurons. This allowed us tobroadly characterize the spontaneous electrophysiological activity of mature neuronalcultures at 56 Days In Vitro, a time point in which the GABA shift has already occurred.
Moreover, we explored the impact of heterogeneity through chemical stimulation. In
particular, the administration of BIC led to an increase in terms of firing and bursting activity
only in the 75E25I configuration, while APV and CNQX caused substantial alterations on bothdynamics and functional connectivity. On the other hand, the electrical stimulation revealed
that the 100E configuration responded reliably, while the 75E25I required additionalparameter tuning for improved responses. Our findings advance the understanding ofdifferent neuronal interactions and their role in the network activity, providing insights forpotential therapeutic interventions in neurological conditions. Overall, our work contributes
to the development of a valuable human in vitro system for investigating physiological andpathological conditions, highlighting the critical role of neuron diversity in neural network
dynamics.
24 . Röck, Ruth

University of Zurich, CH

CELL FATE DETERMINATION OF DIRECTLY REPROGRAMMED MOUSE AND HUMAN KIDNEY
CELLS
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Despite the increasing demand to understand and treat kidney diseases, options formodeling these conditions in vitro remain limited. We are interested in deciphering themolecular programs guiding kidney development and disease and have pioneered the direct
reprogramming of renal tubular cells. By using only four transcription factors we achieve cell
type conversion of fibroblasts to induced renal epithelial cells (iRECs) that aremorphologically and functionally highly similar to primary kidney cells but can be stably
cultivated and bioprinted into tubular structures at near physiological scales. However, theprocess of direct reprogramming isn't fully understood. In-depth analysis, utilizing bulk andsingle-cell RNA sequencing, further characterizes the molecular nature of iRECs concerning
assay stability and cellular heterogeneity. These endeavors aim to refine direct
reprogramming methods, enhancing their effectiveness and reliability for potential
applications in drug testing and disease modeling in the future.
25 . Scherbakov, Dimitri

University of Zurich, CH

ERROR-FREE TRANSLATION AS A NOVEL THERAPEUTIC APPROACH AGAINST AGE-RELATEDNEURODEGENERATIVE DISEASES SUCH AS PARKINSON'S DISEASE
The prevalence of age-related neurodegenerative diseases (NDDs) is increasing worldwide
as the human population ages. Currently, Parkinson's disease (PD) is the second mostcommon NDD, affecting an estimated 7-10 million people worldwide. The increasing
frequency of PD poses a significant challenge to healthcare systems, as PD is typically chronic,
progressive, and associated with high levels of disability and caregiver burden. The
accumulation of protein aggregates, particularly alpha-synuclein, linked with degradation ofproteostatic machinery is thought to play a central role in the development of PD. In turn,protein misfolding/malfunction/aggregation is connected to the accuracy of gene expression
where protein translation is the most error-prone step. Translation errors lead to misfolding
and aggregation of proteins, thus contributing to progression of age-related NDDs such as
PD. We propose to reduce the amount of erroneously translated misfolding-prone proteinsby modulating the work of translation machinery and increasing protein translation fidelity.
Error-free translation reduces the level of intracellular protein misfolding and aggregation
thus representing a promising strategy for treating/preventing the development of NDDs
such as PD. We have identified some genetic and pharmacological interventions decreasingtranslation errors and reducing protein aggregation. A further step of preclinical validation
requires an appropriate experimental model such as PD patient-derived iPSC lines. A large
number of PD patient-derived iPSC lines generated from sporadic patients with different risk
profiles (see the Foundational Data Initiative for PD; www.foundinpd.org) offers a realistic
possibility to analyze potential of increased translation fidelity as therapeutic approach for
PD treatment.
26 . Taborsky, David

University of Zurich, CH

THE ROLE OF TRANSLATIONAL REGULATION IN EARLY DEVELOPMENT AND GASTRULATION
Developmental processes such as gastrulation involve rapid and profound changes in tissue
architecture and cell state. These changes in cell state are the result of alterations in gene
expression of individual cells. Gene expression can be regulated at the level of transcription
and translation, the latter being known as translational control. While transcriptional
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changes have been extensively studied in developmental processes, the potential role oftranslational control in driving the proteome changes required for cellular transformations
are far from understood. We aim to leverage an in vitro organoid model of mammaliangastrulation, the gastruloids, together with ribosome profiling to monitor alteredtranslational efficiencies throughout gastruloid development. Using this approach, we
provide evidence for translational control of various pathways essential for early
development, including components of the Wnt signalling pathway, as well as pervasivetranslational regulation of the translational machinery itself. We then follow up on thistranslational landscape of gastrulation with a single-cell CRISPR screen targeting RNAbinding proteins to uncover the impact of translational control on shaping cell fate and germlayer differentiation in gastruloids.
27 . Tringides, Christina
DESIGNING PHYSIOLOGICALLY-MIMICKINGINTERFACESETH Zurich, CH
PLATFORMSFOR

IMPROVEDNEURAL

Biomaterial scaffolds have emerged as a tool to build 3D cultures of cells which better
resemble biological systems, while advancements in bioelectronics have enabled themodulation of cell proliferation, differentiation, and migration [1]. Here, we first describe aporous conductive hydrogel with the same mechanical modulus and viscoelasticity as neuraltissue [2]. Electrical conductivity is achieved by incorporating low amounts (<0.3% weight)of carbon nanomaterials in an alginate hydrogel matrix, and then freeze-drying to introduce
a highly porous network. The mechanical and electrical properties of the material can betuned to modulate the growth and differentiation of human iPSC neural progenitor cells(NPCs). In addition to forming neurite networks that span the material in 3D, the NPCs candifferentiate into astrocytes and oligodendrocytes. With increasing hydrogel viscoelasticity
and conductivity, we observe the formation of denser neurite networks and a higher degreeof myelination. Application of exogenous electrical stimulation can then be applied to thescaffolds to further promote NPC differentiation. To investigate the functionality of neurite
networks in 3D, we begin by placing a polydimethylsiloxane (PDMS) microstructure on anunderlying multielectrode array (MEA), as previously described [3], to confine the network
growth. We then explore different materials and techniques to integrate hydrogels into the
PDMS microstructures, such that the hydrogel can facilitate human iPSC-derived sensory
neurons to form 3D networks while still confined by the PDMS. This platform is compatible
with various methods to assess neuronal functionality, and can be used to understand theeffect(s) of hydrogel properties on the resulting neuronal networks. Both describedbiomaterial platforms can support the growth of neuronal cells for over 6 weeks, and could
facilitate the development of biohybrid electronic devices to understand neuronaldevelopment and disease.
28 . Vehring, Leah

University of Bern, CH

INVESTIGATING PARACRINE INTERACTIONS IN HIPSC DERIVED IN VITRO MODEL MODEL OF
NEUROVASCULAR UNIT
Pericytes are mural cells supporting brain microvascular endothelial cells (BMECs) at thelevel of brain capillaries and their dysfunction has been implicated in various neurovascular39

disorders. However, only little is known on their direct influence on BMECs in Multiple
Sclerosis (MS) pathogenesis. We therefore want to elucidate the paracrine contribution of
pericytes to the previously observed intrinsic blood brain barrier (BBB) dysfunction andincreased immune cell interaction. To do so, conditioned medium from human iPSC derived
brain pericyte like cells (BPLC) will be analyzed with mass spectrometry to identify possible
candidates that are capable of inducing impaired BBB characteristics in human iPSC-derived
BMECs. This will be assessed applying various assays, such as permeability assay andadhesion assays, as well as junctional protein staining of BMECs. Lastly, these results will becomplemented with the investigation of adhesion molecule expression, namely VCAM-1 andICAM-1, for immune cell interaction. Interestingly, we showed that treatment of BMECs with
BPLC conditioned medium and pro-inflammatory cytokines (TNF-a/IFN-y) increased theexpression of relevant adhesion molecules. Taken together, this study will provide a better
understanding of the role of pericytes in BBB dysfunction in the context of MS.
29 . Vogt, SarahETH Zurich, CH

EXPLORING MORPHOLOGICAL HETEROGENEITY IN HUMAN PLURIPOTENT STEM CELLS WITH
DEEP LEARNING
Induced pluripotent stem cells (iPSCs) exhibit maximal developmental plasticity, enabling
differentiation into various cell types in response to environmental cues, resulting inalterations in their cellular states, including morphology. Investigating these diverse iPSC
morphologies at a high-throughput scale requires efficient characterisation andquantification of morphological variations within high-content imaging data. Deep learning
methods are highly suited for this task, given their efficient pattern recognition capabilities.
In our study, we demonstrate the potential of scDINO (single-cell distillation with no labels),
a self-supervised deep learning approach, to comprehensively characterise the complete
range of cellular morphologies of iPSCs subjected to diverse small molecule perturbations ina fully unbiased fashion. Beyond mere classification, our investigation illustrates how reliably
characterising and quantifying single-cell morphologies opens avenues for mapping bulk
molecular data back to the level of individual cells. This approach holds promise foruncovering intricate relationships between molecular changes and alterations in cellularmorphology, providing deeper insights into iPSC behaviour and differentiation pathways.
30 . Walter, Natalie

University of Zurich, CH

ELUCIDATING THE FUNCTIONAL EFFECTS OF OMEGA-3 FATTY ACIDS AS A TREATMENT IN
ADHD AGAINST INFLAMMATION AND OXIDATIVE STRESS
Attention-deficit hyperactivity disorder (ADHD) is the most frequently reported
neurodevelopmental disorder, with a worldwide prevalence of ca. 5%, affecting children andadolescents. Inflammation and oxidative stress may play a crucial role in ADHD, indicated byaltered serum levels of IL-6 and TNFα cytokines as well as increased reactive oxygen species
discovered in ADHD children. Therefore, the non-pharmacological treatment of omega-3 (ω3) polyunsaturated fatty acids (PUFA), docosahexaenoic acid (DHA) and eicosapentaenoic
acid (EPA), display potential candidates, as they take part in several biological mechanisms,
including regulations of inflammatory processes and attenuating oxidative stress. However,the underlying molecular mechanisms of ω-3 PUFAs involved in ADHD remains unknown.
40Thus, we investigated IL-6 as well as TNFα release and oxidative stress in human induced
pluripotent stem cell (iPSC)- derived forebrain cortical neurons (FCNs) from ADHD patientsand healthy individuals as control after DHA (10μM) and EPA (10μM) treatments.
Investigations of pro-inflammatory cytokines released in FCN supernatants showedtendencies of decreased TNFα secretion in ADHD. These alterations could be increased tocontrol levels after DHA treatment, which may suggest TNFα being crucial for healthy brain
development. Moreover, as mitochondria demonstrate the main source of reactive oxygen
species production we investigated superoxide accumulation derived from mitochondria in
FCNs by conducting MitoSox live imaging. ADHD cell lines were discovered to produce
significantly higher levels of superoxide’s compared to controls (Mann-Whitney, *p=0.0159),
which could be decreased after DHA and EPA treatments. These findings may help
understand the importance of inflammation and oxidative stress more evidently andfurthermore may reveal the functional effects of ω-3 PUFA to be considered as an additional
treatment approach in ADHD in a personalized manner.
31 . Weber, Rebecca Z.

University of Zurich, CH

HUMAN IPSC-DERIVED CELL GRAFTS PROMOTE FUNCTIONAL RECOVERY BY MOLECULAR
INTERACTION WITH STROKE-INJURED BRAIN
Stroke is a leading cause of disability and death due to the brain’s limited ability toregenerate damaged circuits. To date, stroke patients have only limited therapeutic options
and are often left with considerable disabilities. Induced pluripotent stem cell (iPSC)-based
therapies are emerging as a promising therapeutic approach for stroke recovery. In thisstudy, we demonstrate that local transplantation of GMP-compatible iPSC-derived neuralprogenitor cells (NPC) improve long-term recovery-associated brain tissue responses andreduced neurological deficits after cerebral ischemia in mice. We observe long-term graft
survival over the course of five weeks and preferential graft differentiation into matureneurons without signs of pluripotent residuals. Transplantation of NPCs led to a set ofrecovery associated tissue responses including increased vascular sprouting and repair,reduced blood-brain barrier disruption, reduced microglial activation and increased
neurogenesis compared to littermate control animals receiving sham transplantation withphosphate-buffered saline (PBS). Employing deep learning assisted behavior analysis, werevealed improved gait performance and reduced fine-motor deficits. To dissect themolecular graft composition and identify graft-host interactions, we performed single
nucleus profiling of graft and host tissue that identifies graft differentiation towards
preferentially GABAergic-like cells with remaining cells acquiring glutamatergic neuron,
astrocyte, and NPC-like phenotypes. Interactions between graft and host transcriptome
indicate that GABAergic cell grafts are primarily involved in graft-host communication
through regeneration-associated NRXN, NRG, NCAM and SLIT signalling pathways. In
conclusion, our study indicates that transplanted iPSC-derived NPCs differentiate mainly into
GABAergic-like cells contributing to long-term anatomical and functional recovery andidentifies potential mechanisms involved in this process. These results provide a basis for atherapeutic strategy to improve brain regeneration after stroke.

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32 . Yde Ohki, Cristine Marie

University of Zurich, CH

INVOLVEMENT OF THE WNT SIGNALING IN METHYLPHENIDATE (RITALIN) TREATMENT OFATTENTION-DEFICIT HYPERACTIVITY DISORDER
Attention-deficit hyperactivity disorder (ADHD) is a multifactorial neurodevelopmental
disorder that affects over 5% of children and adolescents around the world. However,molecular mechanisms involved in ADHD etiology are still under investigation.
Methylphenidate (MPH), the first-line treatment for ADHD, ameliorates brain maturational
delays in patients. The Wnt signaling pathway might be associated with the ADHD
pathophysiology. We aim to investigate whether MPH modulates Wnt signaling, which might
contribute to clinical improvements observed in patients. ADHD patients who respond to
MPH treatment and matching controls (6-18 years old) were recruited by the Department ofChild and Adolescent Psychiatry and Psychotherapy, University of Zurich. Induced
pluripotent stem cells (iPSCs), iPSC-derived neural stem cells (NSCs) and forebrain corticalneurons (FCNs) were generated. Proteomic analysis via Western Blot was conducted in thesecells and Wnt reporter assays were performed in NSCs to analyze Wnt activity after
treatment with the agonist Wnt3a, the antagonist DKK1, and MPH. Growth rates of iPSCs
and NSCs were also measured via xCELLigence/WST-1. Synaptogenesis in FCNs was assessed
via immunocytochemistry. ADHD NSCs proliferated significantly less than controls for bothxCELLigence (Mann Whitney, *p=0.014) and WST-1 assays (Welch’s t test, *p=0.012). Thiswas slightly improved by MPH 10 nM in a Wnt-dependent manner. Increased active βcatenin and decreased inactive GSK3β (Mann Whitney, *p=0.043 and *p=0.018, respectively)
indicated Wnt signaling alterations in ADHD NSCs, which was confirmed by lower EC50 values
for Wnt3a (Welch’s t test, *p=0.033) and increased IC50 values for DKK1 in reporter assays.
Additionally, Wnt activity seems to be modulated by MPH 10 nM. Synaptic connectivity ishigher in ADHD FCNs (Mann Whitney, *p=0.032). These results might be associated with theclinical outcomes observed in patients and with MPH-induced improvements in this context.
33 . Zanini, Giorgia

University of Genova, IT

EFFECTIVE PARAMETERS FOR ELECTRICAL STIMULATION ON HUMAN IPSCS-DERIVED
NEURONAL NETWORKS COUPLED TO MICRO ELECTRODE ARRAYS
In vitro models of neuronal networks have proven to be a valuable tool for gaining deeper
insights into the intricate mechanisms governing the human brain. Notably, the integrationof human induced pluripotent stem cells (hiPSCs) with micro-electrode arrays (MEAs)
provides a controlled in vitro environment to replicate and analyse both the structural andfunctional elements of the human brain. As neuronal communication relies on the emissionof electrical (and chemical) stimuli, the employment of electrical stimulation becomes crucial
for a comprehensive exploration of neuronal assemblies, to better understand their inherent
electrophysiological dynamics. However, the establishment of standardized stimulation
protocols for cultures derived from hiPSCs is still lacking, thereby hindering the precise
delineation of efficacious parameters to elicit responses. To fill this gap, this study aims toinvestigate the stimulation parameters aiming to induce independent and reliable evoked
responses in human-derived neurons. We utilized biphasic voltage pulses with variable
amplitudes (1.5, 2, 2.5, and 3V) delivered at different frequencies (0.1 and 0.2Hz). As anexperimental model, we exploited glutamatergic (excitatory, E) and GABAergic neurons
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(inhibitory, I) to construct fully excitatory (E:I 100:0) and mixed (E:I 75:25) cortical networks
derived from hiPSCs coupled to MEAs. This approach allowed us not only to optimize
stimulation parameters but also to understand their specific impact on the functional
dynamics of both excitatory and inhibitory elements within the neuronal networks. This
study represents a stepping-stone in the exploration of efficacious stimulation parameters,
thus broadening the electrophysiological activity profiling of neural networks sourced from
hiPSCs.
34 . Ziak, Nicole

University Hospital Zurich, CH

UNDERSTANDING THE SELECTIVE NEUROTOXICITY OF SERINE-PALMITOYLTRANSFERASEMUTATIONS IN MOTOR AND SENSORY NEURONS
Serine–palmitoyltransferase (SPT) is the key enzyme for the de novo synthesis ofsphingolipids (SLs). Mutations in SPT were previously linked to the Hereditary Sensory andAutonomous Neuropathy Type 1 (HSAN1) - a predominantly sensory neuropathy that iscaused by the pathological formation and accumulation of an atypical class of neurotoxic 1deoxySL. Recently, mutations in SPT have also been associated with Amyotrophic Lateral
Sclerosis (ALS), where disease manifests differently. ALS is a progressive, neurodegenerative
disease of the lower and upper motor neurons (MN), characterized by severe muscle
wasting, eventually leading to paralysis and death. An altered homeostatic control leads toan overshooting formation of canonical SL. MN seem to be particularly susceptible to thesechanges whereas sensory neurons (SN) appear to be specifically sensitive to 1-deoxySLs
accumulation, that occurs in HSAN1. In this PhD project, we want to understand how
mutations in the same enzyme –SPT- can lead to such opposing clinical phenotypes. This will
hopefully provide insights into the underlying pathomechanisms of both diseases, but also
in the metabolic differences between SN and MN, opening up the possibility for novel
therapeutic targets. Therefore, we will analyze the lipid profiles in three in vitro models,increasing in complexity. The first two are based on genetic engineering, where neuronaldifferentiation is induced by transcription factor overexpression. These models are called
inducible Neurons (iNeurons). The first experiment uses wild type iNeurons, whereas in thesecond one, ALS and HSAN1 specific mutations are introduced by CRIPSR-Cas9. The thirdmodel, which is the most physiological one, uses patient derived iPSCs. These iPSCs will bedifferentiated according a small-molecule approach into MN and SN. The lipid profile ofmature neurons and how it changes during the differentiation and the effect of perturbing
the SL pathway will be assessed in all three models.
35 . Zujur, Denise

Kyoto University, JP

GENERATION OF IPSC-DERIVED CHONDROGENIC SPHEROIDS VIA NEURAL CREST CELLINDUCTION FOR CARTILAGE REPAIR AND BIOFABRICATION
Articular cartilage is a highly specialized connective tissue providing a lubricated surface tobone joints. Unlike most tissues, articular cartilage does not have blood vessels, nerves, or
lymphatics, resulting in limited self-healing capacity. To date, there is no effective longlasting treatment to repair articular cartilage. In terms of regenerative medicine approach,primary chondrocytes and mesenchymal stem cells (MSCs) are the most common cellsources used in clinical settings for repairing articular defects. However, both cell types
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possess limitations such as dedifferentiation, donor morbidity and limited expansion. We
developed a stepwise differentiation method to generate cartilage spheroids from iPSCs via
neural crest cell (iNCC) induction. iNCCs are further differentiated into iMSCs andchondrocytes. The cell strategy uses a combination of growth factors and small molecule
inducers to produce controlled-size spheroids and enhanced cartilage extracellular matrix
production with no sign of dedifferentiation, fibrotic cartilage formation or hypertrophy. In
particular, we demonstrate that the use of a thienoindazole derivative, TD-198946,
synergistically enhances chondrogenesis of iMSCs derived from iNCCs. These findings may
provide a novel cell source for stem cell-based cartilage therapy. In addition, as thechondrogenic spheroids also have the potential to fuse within few hours, we are using themas building blocks for biofabrication of larger cartilage tissue using technologies such as the
bioprinting Kenzan method.

Please don’t print me.

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