In a study published in the peer-reviewed journal Cell Stem Cell, investigators detail how they created “ALS on a chip” and the clues the specialized laboratory chip has already produced about nongenetic causes of the disease, also known as Lou Gehrig’s disease.

The work builds on previous studies where adult cells from ALS patients were reverted into stem cells. The cells were then pushed forward to produce motor neurons, which die in the disease, causing progressive loss of the ability to move, speak, eat and breathe.

In this study, the motor neurons from ALS patients were seeded into the top channels of microengineered chips. Cells that make up the blood-brain barrier were seeded into the bottom channels of the chips. The two channels are connected through a porous membrane that allows investigators to flow fluids through the chips in order to mimic blood flow.

Investigators created a second group of the specialized chips using cells from individuals who did not have ALS, then used advanced technologies to analyze more than 10,000 genes in the motor neurons in both groups of chips.

“In our early work, we couldn’t detect many differences between the motor neurons of patients with ALS and those from healthy individuals,” said Clive Svendsen, PhD, executive director of the Board of Governors Regenerative Medicine Institute at Cedars-Sinai and senior author of the study. “But those studies employed traditional lab culture that is static like a pond. In the body, blood vessels provide constant fluid flow to bring in nutrients and take away waste—and may even provide other types of support to motor neurons.”

In the specialized chips, the motor neurons matured more completely than they would in a static dish, and investigators could detect distinct differences in the cells from patients with ALS.

“We were intrigued to find that signaling for glutamate, a chemical that sends excitatory messages between neurons, was altered in the ALS motor neurons,” Svendsen said. “Excessive release of glutamate has long been considered a possible cause of ALS, and one of the few drugs approved to treat the disease targets this neurotransmitter. The changes we found don’t seem to cause any issues for the motor neurons when they are young, but over many years it is possible that this increased glutamate signaling may be part of why motor neurons die in ALS.”

Svendsen said that while these results are exciting, the team’s next task is to determine whether this increased glutamate signaling directly leads to the dysfunction or death of the cells. He also noted that glutamate is likely only one piece in a much larger puzzle that underlies the cause of ALS.

“These models allow us to better understand the very earliest stages of the disease process,” Svendsen said. “We haven’t connected all the dots yet, but based on these findings we have a model that will allow us to test our theories. If we can show that glutamate signaling eventually makes the ALS motor neurons sick, for instance, we can apply drugs to the blood vessel side of the chip to mimic a clinical trial. Those experiments are underway.”

Additional authors include Deepti Lall, Michael Workman, Samuel Sances, Briana N. Ondatje, Shaughn Bell, George Lawless, Amanda Woodbury, Dylan West, Amanda Meyer, Andrea Matlock, Vineet Vaibhav, and Jennifer E. Van Eyk.

Funding: This work was supported by NIH-NCATS grants 1UG3NS105703-01 and 1UG3TR003264-01, and funding from The ALS Association.

Cedars-Sinai owns a minority stock interest in Emulate, the company that produces the study’s microfluidic Organ-Chips. An officer of Cedars-Sinai also serves on Emulate’s Board of Directors. C.N.S serves on the advisory board of Cell Stem Cell.

Cedars-Sinai Health Sciences University is advancing groundbreaking research and educating future leaders in medicine, biomedical sciences and allied health sciences. Learn more about the university.

, hosted by Cedars-Sinai on Oct. 25.

“This meeting isn’t just for scientists,” said Michael I. Lewis, MD, program director of The Alpha Clinic at Cedars-Sinai and professor of Medicine. “It is for clinicians, patients and members of the public who are interested in learning more about cell and gene therapies, emerging technologies, innovative clinical trial designs and issues surrounding accessibility to these therapies for patients, such as financial barriers.”  

The symposium, sponsored by the California Institute for Regenerative Medicine (CIRM), will take place from 9 a.m.-5 p.m. at the Cedars-Sinai Silver Screen Theater in the Pacific Design Center. Virtual registration is also available.

Scientists from all nine California Alpha Clinics are set to speak. Along with Cedars-Sinai, institutions with alpha clinics include University of California, San Diego; University of California, Los Angeles; University of Southern California and Children’s Hospital Los Angeles; Stanford University; University of California, Irvine; University of California, Davis; City of Hope; and University of California, San Francisco.

Directors of several Alpha Clinics will moderate panel discussions, and directors of the Alpha Clinics at UC Irvine, UC San Diego, University of Southern California and UC Davis will participate in a discussion of Alpha Clinic operations. Daniela A. Bota, MD, PhD, director of the UC Irvine Alpha Clinic, will talk about chimeric antigen receptor (CAR) therapies, and Mark Walters, MD, UC San Francisco Alpha Clinic director, will discuss current gene therapies, future opportunities and barriers to treatment. 

Lewis expects speakers to highlight the importance of diversity in clinical trials, promote collaboration among researchers and showcase advances in cell and gene therapies.

“The symposium will look to the current status of the field and, more importantly, to the future, as new approaches and therapies are likely to rapidly expand,” said Clive Svendsen, PhD, co-associate project director of Cedars-Sinai's Alpha Clinic and executive director of the Board of Governors Regenerative Medicine Institute at Cedars-Sinai. Svendsen recently co-authored a review paper published in Nature Medicine about stem cell-based therapies for neurological conditions.

The review paper describes stem cell-based treatments for a number of neurological disorders, including amyotrophic lateral sclerosis, also known as ALS. The condition affects nerve cells in the brain and spinal cord that control movement. ALS eventually causes loss of the ability to move, speak, eat and breathe. At this time there is no cure.

The Cedars-Sinai Alpha Clinic is conducting a trial of a therapy, developed by Svendsen and his team, using specially engineered cells to protect damaged motor neurons. To date, six patients have been treated, with no major complications.

Cedars-Sinai’s Alpha Clinic and others throughout California are developing stem-cell based therapies for a variety of conditions: 

• Cedars-Sinai is studying a therapy for retinitis pigmentosa, which breaks down cells in the light-sensitive layer at the back of the eye, the retina, leading to vision loss. The therapeutic cells in the trial release proteins that help protect retinal cells from damage.
• Cedars-Sinai investigators also are testing a treatment for pulmonary arterial hypertension, a rare condition in which elevated blood pressure in lung blood vessels can damage the heart. The initial study was led by Lewis, director of Respiratory Care Services at Cedars-Sinai, and Eduardo Marbán, MD, PhD, executive director of the Smidt Heart Institute and associate project director of The Alpha Clinic. Investigators concluded that infusions of specialized cell products derived from heart tissue seemed to improve patients’ heart and lung health.
• Alpha Clinics throughout California are studying stem cell-based therapies for several other conditions—including movement disorders such as Parkinson’s disease and Huntington’s disease; Alzheimer’s disease; stroke; epilepsy; spinal cord injury; and multiple sclerosis.

“The Alpha Clinic network is dedicated to accelerating the work of institutions in California, such as Cedars-Sinai, that are developing and testing cell and gene therapies,” Marbán said. “It is a highly collaborative group, and we share resources including our expertise, our infrastructure and our experience with cell delivery to help each other.”

Follow Cedars-Sinai Academic Medicine on LinkedIn for more on the latest basic science and clinical research from Cedars-Sinai.

and professor of Medicine and Biomedical Sciences at Cedars-Sinai, is developing new treatments and models for amyotrophic lateral sclerosis (ALS) using stem cells.

ALS progressively destroys nerve cells in the brain and spinal cord, causing people to lose muscle movement required for walking, talking, swallowing and breathing. Currently, the disease is irreversible and has no cure, but a potential therapy that combines gene and stem cell technologies is progressing well, according to Svendsen.

Svendsen has been selected as an inaugural recipient of a grant from the Tambourine ALS Breakthrough Research Fund, funded by Tambourine and administered in partnership with the Milken Institute. The grant will fund research that combines advanced stem cell modeling with artificial intelligence to learn more about the cause of ALS. Svendsen sat down with the Cedars-Sinai Newsroom to discuss his ALS research and new projects that the grant will help fund.

What is unique about the ALS therapy you are developing?

It uses a protein called GDNF (glial cell line-derived neurotrophic factor), which is a nutrient that brain cells can use to mature and survive. It promotes the growth of axons, the threadlike structures brain cells use to communicate, and also helps damaged cells rejuvenate.  

In ALS, motor neurons are dying. And there is evidence that GDNF can help protect them. The problem is that GDNF is a large protein and it cannot pass through the barrier that protects the brain by preventing potentially harmful substances from entering it.

So how do you get GDNF into the brain and spinal cord?

We've developed a clever technique, like a Trojan horse. We've engineered a human stem cell called a neural progenitor to release GDNF, and we're transplanting those cells into the spinal cords and brains of patients with ALS. The idea is that the progenitor cells mature into healthy support cells and release GDNF that will help heal diseased motor neurons. 

How are clinical trials of the therapy going?

Our first trials are to ensure the therapy is safe for patients. We have completed a Phase I/IIa clinical trial to treat the lower motor neuron, which controls leg movement, in 18 patients with ALS. We had good safety data in every one of those patients. Those results were published in Nature Medicine.

We only put the cells on one side of each patient’s spinal cord, so we could compare the treated leg to the nontreated leg. While the focus of this initial study was safety, we did see a trend toward improvement in the treated limb after 12 months, even though the patients were at an advanced disease stage. We plan to perform an additional trial with patients who are earlier in the disease process and with optimized cell targeting, and hope to significantly improve limb function.

As the upper motor neuron in the brain is also affected in ALS, we’ve started an additional safety trial where we will treat the upper motor neurons, specifically those that control hand movement.  We've treated five patients and the trial is still open. Our next step will be to treat both the upper and lower motor neurons, and at that point, we will also measure the effectiveness of the therapy. If our technique is working, we should see a slowing of disease progression in the treated hand and leg. Research Project Advisor Pablo Avalos, MD; Adam Mamelak, MD, director of the Functional Neurosurgery Program at Cedars-Sinai; and Richard Lewis, MD, director of the Electromyography Laboratory at Cedars-Sinai, are key contributors to this work.

What will the Tambourine grant fund?

With the help of the Cedars-Sinai Biomanufacturing Center, as part of a large consortium called Answer ALS, we have generated induced pluripotent stem cells from 1,000 patients with ALS. We have full clinical records and genomic sequencing for these patients. Induced pluripotent stem cells are adult human cells taken back in time to a stage where they are capable of developing into any cell in the body. We’re then differentiating those cells into motor neurons that are known to die in ALS patients.

In collaboration with MIT [Massachusetts Institute of Technology], we will use the Tambourine grant to fund an advanced imaging project, called cell painting, on these motor neurons. Michael Workman, a project scientist in my laboratory, has been a key contributor to these studies. Up to six fluorescent dyes are used to label different components of the cells. With the help of an advanced, high-powered microscope and massive cloud computing, each individual neuron will be analyzed. 

We will use artificial intelligence to attempt to distinguish ALS patients from healthy patients, and then look for molecular “signatures” within ALS patients that define subgroups. This data can be used to develop new targeted drugs to treat specific subgroups of ALS—a truly personalized medicine approach leveraging the latest medical technology. This could be a big boost for clinical trials and drug development.

Read more on the Cedars-Sinai Blog: Regenerative Medicine: A New Path for ALS Treatment