A miniature heart in a petri dish: Organoid emulates development of the human heart

A team at the Technical University of Munich (TUM) has induced stem cells to emulate the development of the human heart. The result is a sort of "mini-heart" known as an organoid. It will permit the study of the earliest development phase of our heart and facilitate research on diseases.

The human heart starts forming approximately three weeks after conception. This places the early phase of heart development in a time when women are often still unaware of their pregnancy. That is one reason why we still have little knowledge of many details of how the heart is formed. Findings from animal studies are not fully transferable to humans. An organoid developed at TUM could prove helpful to researchers.

A ball of 35,000 cells

The team working with Alessandra Moretti, Professor of Regenerative Medicine in Cardiovascular Disease, has developed a method for making a sort of "mini-heart" using pluripotent stem cells. Around 35,000 cells are spun into a sphere in a centrifuge. Over a period of several weeks, different signaling molecules are added to the cell culture under a fixed protocol. "In this way, we mimic the signaling pathways in the body that control the developmental program for the heart," explains Alessandra Moretti. The group has now published its work in the journal Nature Biotechnology.

First-ever "epicardioids"

The resulting organoids are about half a millimeter in diameter. Although they do not pump blood, they can be stimulated electrically and are capable of contracting like human heart chambers. Prof. Moretti and her team are the first researchers in the world to successfully create an organoid containing both heart muscle cells (cardiomyocytes) and cells of the outer layer of the heart wall (epicardium). In the young history of heart organoids -- the first were described in 2021 -- researchers had previously created only organoids with cardiomyocytes and cells from the inner layer of the heart wall (endocardium).

"To understand how the heart is formed, epicardium cells are decisive," says Dr. Anna Meier, first author of the study. "Other cell types in the heart, for example in connecting tissues and blood vessels, are formed from these cells. The epicardium also plays a very important role in forming the heart chambers." The team has appropriately named the new organoids "epicardioids."

New cell type discovered

Along with the method for producing the organoids, the team has reported its first new discoveries. Through the analysis of individual cells they have determined that precursor cells of a type only recently discovered in mice are formed around the seventh day of the development of the organoid. The epicardium is formed from these cells. "We assume that these cells also exist in the human body -- if only for a few days," says Prof. Moretti.

These insights may also offer clues as to why the fetal heart can repair itself, a capability almost entirely absent in the heart of an adult human. This knowledge could help to find new treatment methods for heart attacks and other conditions.

Producing "personalized organoids"

The team also showed that the organoids can be used to investigate the illnesses of individual patients. Using pluripotent stem cells from a patient suffering from Noonan syndrome, the researchers produced organoids that emulated characteristics of the condition in a Petri dish. Over the coming months the team plans to use comparable personalized organoids to investigate other congenital heart defects.

With the possibility of emulating heart conditions in organoids, drugs could be tested directly on them in the future. "It is conceivable that such tests could reduce the need for animal experiments when developing drugs," says Alessandra Moretti.

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Human brain organoids implanted into mouse cortex respond to visual stimuli for first time

A team of engineers and neuroscientists has demonstrated for the first time that human brain organoids implanted in mice have established functional connectivity to the animals' cortex and responded to external sensory stimuli. The implanted organoids reacted to visual stimuli in the same way as surrounding tissues, an observation that researchers were able to make in real time over several months thanks to an innovative experimental setup that combines transparent graphene microelectrode arrays and two-photon imaging.

The team, led by Duygu Kuzum, a faculty member in the University of California San Diego Department of Electrical and Computer Engineering, details their findings in the Dec. 26 issue of the journal Nature Communications. Kuzum's team collaborated with researchers from Anna Devor's lab at Boston University; Alysson R. Muotri's lab at UC San Diego; and Fred H. Gage's lab at the Salk Institute.

Human cortical organoids are derived from human induced pluripotent stem cells, which are usually derived themselves from skin cells. These brain organoids have recently emerged as promising models to study the development of the human brain, as well as a range of neurological conditions.

But until now, no research team had been able to demonstrate that human brain organoids implanted in the mouse cortex were able to share the same functional properties and react to stimuli in the same way. This is because the technologies used to record brain function are limited, and are generally unable to record activity that lasts just a few milliseconds.

The UC San Diego-led team was able to solve this problem by developing experiments that combine microelectrode arrays made from transparent graphene, and two-photon imaging, a microscopy technique that can image living tissue up to one millimeter in thickness.

"No other study has been able to record optically and electrically at the same time," said Madison Wilson, the paper's first author and a Ph.D. student in Kuzum's research group at UC San Diego. "Our experiments reveal that visual stimuli evoke electrophysiological responses in the organoids, matching the responses from the surrounding cortex."

The researchers hope that this combination of innovative neural recording technologies to study organoids will serve as a unique platform to comprehensively evaluate organoids as models for brain development and disease, and investigate their use as neural prosthetics to restore function to lost, degenerated or damaged brain regions.

"This experimental setup opens up unprecedented opportunities for investigations of human neural network-level dysfunctions underlying developmental brain diseases," said Kuzum.

Kuzum's lab first developed the transparent graphene electrodes in 2014 and has been advancing the technology since then. The researchers used platinum nanoparticles to lower the impedance of graphene electrodes by 100 times while keeping them transparent. The low-impedance graphene electrodes are able to record and image neuronal activity at both the macroscale and single cell levels.

By placing an array of these electrodes on top of the transplanted organoids, researchers were able to record neural activity electrically from both the implanted organoid and the surrounding host cortex in real time. Using two-photon imaging, they also observed that mouse blood vessels grew into the organoid providing necessary nutrients and oxygen to the implant.

Researchers applied a visual stimulus-an optical white light LED-to the mice with implanted organoids, while the mice were under two-photon microscopy. They observed electrical activity in the electrode channels above the organoids showing that the organoids were reacting to the stimulus in the same way as surrounding tissue. The electrical activity propagated from the area closest to the visual cortex in the implanted organoids area through functional connections. In addition, their low noise transparent graphene electrode technology enabled electrical recording of spiking activity from the organoid and the surrounding mouse cortex. Graphene recordings showed increases in the power of gamma oscillations and phase locking of spikes from organoids to slow oscillations from mouse visual cortex. These findings suggest that the organoids had established synaptic connections with surrounding cortex tissue three weeks after implantation, and received functional input from the mouse brain. Researchers continued these chronic multimodal experiments for eleven weeks and showed functional and morphological integration of implanted human brain organoids with the host mice cortex.

Next steps include longer experiments involving neurological disease models, as well as incorporating calcium imaging in the experimental set up to visualize spiking activity in organoid neurons. Other methods could also be used to trace axonal projections between organoid and mouse cortex.

"We envision that, further along the road, this combination of stem cells and neurorecording technologies will be used for modeling disease under physiological conditions; examining candidate treatments on patient-specific organoids; and evaluating organoids' potential to restore specific lost, degenerated or damaged brain regions," Kuzum said.

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Organoids reveal how SARS-CoV-2 damages brain cells -- and a potential treatment

Using human brain organoids, an international team of researchers, led by scientists at University of California San Diego School of Medicine and Sanford Consortium, has shown how the SARS-CoV-2 virus that causes COVID-19 infects cortical neurons and specifically destroys their synapses -- the connections between brain cells that allow them to communicate with each other.

The findings, published in the November 3, 2022 issue of PLOS Biology, also report that the antiviral drug sofosbuvir, already an approved treatment for hepatitis C, effectively inhibited SARS-CoV-2 replication and reversed neuronal alterations in infected brain organoids.

"Vaccines and emerging treatments have reduced the health consequences of COVID-19 in most patients," said senior study author Alysson R. Muotri, PhD, professor in departments of Pediatrics and Cellular and Molecular at UC San Diego School of Medicine. "But the phenomenon of Long COVID, characterized by persisting symptoms that include neurological impairment, remains poorly understood and without any specific remedy.

"This work helps explain some of the neurological symptoms of COVID-19 and, more importantly, it suggests that an FDA-approved antiviral drug might be repurposed to restore infected brain cells to health and address long-term neurological outcomes of COVID-19."

Though primarily considered to be a respiratory disease, COVID-19 can cause temporary or long-lasting neurological symptoms in some patients, ranging from loss of taste and smell, impaired concentration (brain fog), and psychological effects such as depression to stroke, epilepsy, and encephalopathy (a change in brain function or structure).

With evidence accumulating that the SARS-CoV-2 virus can infect and alter brain cells (including in developing fetuses), the research team focused on using organoids -- self-organizing, three-dimensional tissues derived from cultured stem cells that can mimic some organ functions.

Researchers exposed the brain organoids to SARS-CoV-2, observed viral infection and replication and noted that the virus rapidly decreased the number of excitatory synapses in neurons within seven days post-infection. Excitatory synapses increase the firing action potential of a neuron, while their counterparts, called inhibitory synapses, decrease that potential.

However, when infected organoids were treated with sofosbuvir, viral replication was inhibited, and observed neurological impairments rescued or restored. The findings echo earlier computational models that suggested sofosbuvir could be a treatment and previous research by Muotri and colleagues that found sofosbuvir effectively protected and rescued neural cells infected by the Zika virus.

"The bottom line is that sofosbuvir appears to have the potential to arrest or prevent the development of neurological symptoms in COVID-19 patients," Muotri said. "And because it has been shown to present no safety concerns in pregnant women, it might also be an option for preventing SARS-CoV-2 transmission to their unborn children.

"Further studies and clinical trials are needed, of course, but these findings offer a path forward for treating a condition (Long COVID) that has so far stymied remedy for millions of people wordwide."

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