Lab-on-Chip Devices For Analysis of Extracellular Vesicles and Their Contents From Cerebral Organoid Tissue
Holger Schmidt, Narinder Kapany Professor of Electrical Engineering, University of California-Santa Cruz, United States of America

Cultured organoids have emerged as important test beds for studying cell development and disease. Sub-micron sized extracellular vesicles (EVs), including exosomes, secreted by these organoids provide an ideal opportunity for assessing and continuously monitoring organoid status on the molecular level. I will describe the development of integrated lab-on-chip devices for real-time analysis of EVs produced in a custom 3D cerebral organoid tissue culture platform. This includes the detection of individual exosomes as well as their molecular biomarker cargo using single particle fluorescence and trapping-assisted nanopore sensing. I will also discuss the direct incorporation of this electro-optofluidic lab-on-chip within the tissue culture platform.

Engineering Organotypic Disease On-a-Chip Models; Harnessing Innovations in Microfluidics, Biomaterials and Single-Cell Resolution Analysis
Mehdi Nikkhah, Associate Professor of Bioengineering, Arizona State University, United States of America

Applications of Human Brain Organogenesis
Alysson Muotri, Professor, Director of the Stem Cell Program, University of California-San Diego, United States of America

Structural and transcriptional changes during early brain maturation follow fixed developmental programs defined by genetics. However, whether this is true for functional network activity remains unknown, primarily due to experimental inaccessibility of the initial stages of the living human brain. We developed cortical organoids that spontaneously display periodic and regular oscillatory network events that are dependent on glutamatergic and GABAergic signaling. These nested oscillations exhibit cross-frequency coupling, proposed to coordinate neuronal computation and communication. As evidence of potential network maturation, oscillatory activity subsequently transitioned to more spatiotemporally irregular patterns, capturing features observed in preterm human electroencephalography (EEG). These results show that the development of structured network activity in the human neocortex may follow stable genetic programming, even in the absence of external or subcortical inputs. Our approach provides novel opportunities for investigating and manipulating the role of network activity in the developing human cortex. Applications for neurodevelopmental disorders, brain evolution and space exploration will be discussed.

Organoids with Reversed Biopolarity (ORBs): SARS-CoV-2 Drug Testing and Breast Cancer Progression
Shuichi Takayama, Professor, Georgia Research Alliance Eminent Scholar, and Price Gilbert, Jr. Chair in Regenerative Engineering and Medicine Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology & Emory University School of Medicine, United States of America

This presentation will describe production of sub-millimeter diameter, consistent size and shape, lung, kidney, and mammary organoids that have an inverted, apical-out geometry. These Organoids with Reversed Biopolarity (ORBs) are used in 384 well plates in a single-organoid-per-well format to test drugs and disease physiology. The airway ORBs are infected with high yields with multiple SARS-CoV-2 strains with Omicron variant showing highest viral replication and Delta giving the most inflammatory response. The ORBs also predict anti-viral drug efficacy correctly where conventional 2D cultures give false signals. Early-stage breast cancer progression model studies will also be discussed.

Leveraging Patient-derived Tumor Organoids For Precision Medicine
Alice Soragni, Assistant Professor, University of California Los Angeles, United States of America

Patient-derived tumor organoids (PDTOs) are representative of the histopathology and physiological behavior of tumors. They are clinically relevant, tractable ex vivo models that can be quickly established from tumor biopsies and surgical samples. There is increasing interest in leveraging tumor organoids for drug development and personalized medicine applications for their ability to maintain principal features of the tumor they originate from, including drug response. PDTOs are particularly important to model rare tumors which often completely lack experimental models. We routinely establish organoids from a spectrum of tumors, including rare ovarian and peritoneal cancers (Phan et al, Communications Biology, 2019), benign tumors such as cutaneous neurofibromas (Nguyen et al, Cell Reports Methods, 2024), indolent bone cancers (Al Shihabi et al, Science Advances, 2022) and aggressive sarcomas (Al Shihabi et al, in press, 2024). We have developed a unique organoid screening platform that uses a modified geometry to seed or bioprint cells in a robust, high throughput and automation-compatible format that bypasses the need for any cell sorting, passaging or in vitro expansion. Our short screening timeline, with results available within one week from surgery, is compatible with therapeutic decision making. Overall, we have been able to identify unique drug responses to both fast and slow-growing cancers, including for tumors that are recalcitrant or impossible to grow as patient-derived xenografts. In a pilot study of PDTOs established from over 120 sarcomas, a family of rare tumors arising from bone or soft tissue, we demonstrated how PDTO drug screening provides sensitivity information that correlates with clinical features yielding actionable information for treatment guidance (Al Shihabi et al, in press, 2024). Importantly, sarcoma organoid responses complemented genetic sequencing and mirrored patient outcomes, leading to the launch of a clinical trial to test PDTO use in osteosarcoma (An Organoid-based Functional Precision Medicine Trial in Osteosarcoma: PREMOST, NCT06064682).

Title to be Confirmed.
Hervé Tiriac, Assistant Researcher, University of California-San Diego, United States of America