Studying human brain disorders is one of the most challenging fields in biomedical research. Besides the fact that the brain is incredibly complex, it’s just plain difficult to peer into it.
It’s neither practical nor ethical to access the cells of the adult brain. Patrick J. Lynch, medical illustrator; C. Carl Jaffe, MD, cardiologist.
For one thing, it’s not practical, let alone ethical, to drill into an affected person’s skull and collect brain cells to learn about their disease. Another issue is that the faulty cellular and molecular events that cause brain disorders are, in many cases, thought to trace back to fetal brain development before a person is even born. So, just like a detective looking for evidence at the scene of a crime, neurobiologists can only piece together clues after the disease has already occurred.
The good news is these limitations are falling away thanks to human induced pluripotent stem cells (iPSCs), which are generated from an individual’s easily accessible skin cells. Last week’s CIRM-funded research report out of Stanford University is a great example of how this method is providing new human brain insights.
Using brain tissue grown from patient-derived iPSCs, Dr. Sergiu Pasca and his team recreated the types of nerve cell circuits that form during the late stages of pregnancy in the fetal cerebral cortex, the outer layer of the brain that is responsible for functions including memory, language and emotion. With this system, they observed irregularities in the assembly of brain circuitry that provide new insights into the cellular and molecular causes of neuropsychiatric disorders like autism.
Recreating interactions between different regions of the development from skin-derived iPSCs
Image: Sergui Pasca Lab, Stanford University
Holy Brain Balls! Recreating the regions of our brain with skin cells
Two years ago, Pasca’s group figured out a way grow iPSCs into tiny, three-dimensional balls of cells that mimic the anatomy of the cerebral cortex. The team showed that these brain spheres contain the expected type of nerve cells, or neurons, as well as other cells that support neuron function.
Still, the complete formation of the cortex’s neuron circuits requires connections with another type of neuron that lies in a separate region of the brain. These other neurons travel large distances in a developing fetus’ brain over several months to reach the cortical cortex. Once in place, these migrating neurons have an inhibitory role and can block the cortical cortex nerve signals. Turning off a nerve signal is just as important as turning one on. In fact, imbalances in these opposing on and off nerve signals are suspected to play a role in epilepsy and autism.
So, in the current Nature study, Pasca’s team devised two different iPSC-derived brain sphere recipes: one that mimics the neurons found in the cortical cortex and another that mimics the region that contains the inhibitory neurons. Then the researchers placed the two types of spheres in the same lab dish and within three days, they spontaneously fused together.
Under video microscopy, the migration of the inhibitory neurons into the cortical cortex was observed. In cells derived from healthy donors, the migration pattern of inhibitory neurons looked like a herky-jerkey car being driven by a student driver: the neurons would move toward the cortical cortex sphere but suddenly stop for a while and then start their migration again.
“We’ve never been able to recapitulate these human-brain developmental events in a dish before,” said Pasca in a press release, “the process happens in the second half of pregnancy, so viewing it live is challenging. Our method lets us see the entire movie, not just snapshots.”
New insights into Timothy Syndrome may also uncover molecular basis of autism
To study the migration of the inhibitory neurons in the context of a neuropsychiatric disease, iPSCs were generated from skin samples of patients with Timothy syndrome, a rare genetic disease which carries a wide-range of symptoms including autism as well as heart defects.
The formation of brain spheres from the patient-derived iPSCs proceeded normally. But the next part of the experiment revealed an abnormal migration pattern of the neurons. The microscopy movies showed that the start and stop behavior of neurons happened more frequently but the speed of the migration slowed. The fascinating video below shows the differences in the migration patterns of a healthy (top) versus a Timothy sydrome-derived neuron (bottom). The end result was a disruption of the typically well-organized neuron circuitry.
Now, the gene that’s mutated in Timothy Syndrome is responsible for the production of a protein that helps shuttle calcium in and out of neurons. The flow of calcium is critical for many cell functions and adding drugs that slow down this calcium flux restored the migration pattern of the neurons. So, the researchers can now zero in their studies on this direct link between the disease-causing mutation and a specific breakdown in neuron function.
The exciting possibility is that, because this system is generated from a patient’s skin cells, experiments could be run to precisely understand each individual’s neuropsychiatric disorder. Pasca is eager to see what new insights lie ahead:
“Our method of assembling and carefully characterizing neuronal circuits in a dish is opening up new windows through which we can view the normal development of the fetal human brain. More importantly, it will help us see how this goes awry in individual patients.”