Confocal microscopy images show mesenchymal stem cells (green) captured within nanovials (pink). Nanovial technology was developed by Dino Di Carlo and colleagues at UCLA. Credit: Shreya Udani/UCLA
University of California Los Angeles Stem cell scientists have uncovered surprising genetic instructions for promoting protein secretion, with major implications for biotechnology and cell therapy.
Mesenchymal stem cells present in the bone marrow secrete therapeutic proteins that may help regenerate damaged tissue.
The UCLA study examining these cells challenges conventional understanding of what genetic instructions drive the release of these therapeutic proteins.
The discovery could help advance both regenerative medicine research and the laboratory production of biological therapeutics already in use.
Expanding the possibilities of antibody-based medicineToday, drugs based on antibodies (proteins that fight infection and disease) are prescribed for everything from cancer to disease. COVID-19 (new coronavirus infection) For high cholesterol. Antibody drugs are supplied by genetically engineered cells that act as small protein-producing factories in the lab.
Meanwhile, researchers are targeting cancer, internal organ damage, and many other diseases with a new strategy that involves transplanting similarly engineered cells directly into patients.
These biotechnological applications rely on the principle of causing cellular changes. DNA When a cell produces more genetic instructions to make a particular protein, it releases more of that protein.
Challenging established biological principlesBut a groundbreaking study from UCLA challenges this long-held belief, at least when it comes to certain types of stem cells.
The researchers looked at mesenchymal stem cells, which reside in the bone marrow and can self-renew and grow into bone, fat, and muscle cells. Mesenchymal cells secrete a protein growth factor called VEGF-A. Scientists believe this may play a role in blood vessel regeneration, repairing damage caused by heart attacks, kidney damage, arterial disease in the extremities, and other diseases.
Amazing discoveries in stem cell researchWhen the researchers compared the amount of VEGF-A released by each mesenchymal cell to the expression of the gene encoding VEGF-A in the same cells, the results were surprising. There was only a weak correlation between gene expression and actual growth factor secretion. Scientists have identified other genes that better correlate with growth factor secretion, including genes that code for proteins on the surface of some stem cells. The research team isolated stem cells with the protein on their surface, cultured a population that secreted large amounts of VEGF-A, and continued to secrete it even after several days.
Biotechnology and its impact on medicineThe findings were published Dec. 11 in the journal natural nanotechnologyco-author Dino Di Carlo said, suggesting that fundamental assumptions in biology and biotechnology may be worth reconsidering. UCLA Samueli School of Engineering.
“The central dogma is that there are instructions in DNA, and these instructions are transcribed. RNAThe RNA is then translated into protein,” said Di Carlo, who is also a member of UCLA. California Nanosystems Institute and Eli and Edythe Regional Center for Regenerative Medicine and Stem Cell Research. “Based on this, many scientists assumed that if you had more RNA, you would get more protein, and more protein would be released from the cell. I had doubts.”
It seems inconceivable that when a gene is expressed at a higher level, there is more secretion of the corresponding protein. We found a clear example where this does not occur, and many new questions arise.” Ta.
“The results could help make the production of antibody-based therapeutics more efficient and define new, more effective cell therapies. Knowing the right genetic switches to flip could enable the manipulation and selection of highly productive cells to create or deliver therapeutics.
Breakthrough in single cell analysisThe UCLA study was conducted using standard laboratory equipment enhanced with technology invented by Di Carlo and his colleagues. Nanovials, microscopic bowl-shaped hydrogel containers, each capturing a single cell and its secretions. By leveraging a new analytical method using nanovials, scientists were able to measure the amount of VEGF-A released by each of 10,000 mesenchymal stem cells compared to tens of thousands of genes expressed by that same cell. I was able to link it to the mapped atlas.
“The ability to link protein secretion to gene expression at the single-cell level holds great promise for the fields of life science research and therapeutic development,” said David, a member of the Broad Stem Cell Research Group and a professor of biology at the University of California, Los Angeles. said chemistry professor Kathryn Plath. Center and co-corresponding author of the study. “Without that, we would not have been able to reach the unexpected results found in this study. Now we have learned something new about the mechanisms that underpin the fundamental processes of life, and we have We have an incredible opportunity to leverage this to improve human health.”
A new path in therapeutic drug developmentAlthough activation of genetic instructions for VEGF-A showed little correlation with protein release, the researchers identified a cluster of 153 genes with strong associations with VEGF-A secretion. Many of them are known for their functions in blood vessel development and wound healing. For others, their functionality is currently unknown.
One of the top matches encodes the cell surface protein IL13RA2, but its purpose is poorly understood. Its outer location made it easy for scientists to use it as a marker and separate those cells from other cells. Cells with IL13RA2 showed 30% more VEGF-A secretion than cells lacking the marker.
In a similar experiment, the researchers kept isolated cells in culture for six days. At the end of that period, cells with the marker secreted 60% more VEGF-A compared to cells without the marker.
Potential impact on clinical applicationsMesenchymal stem cell-based therapies have shown promise in laboratory studies, but many of these new options are safe but not effective in clinical trials with human participants. It is shown that there is no. Her ability to use IL13RA2 to sort VEGF-A-rich cells could help change this trend.
“Identifying the subpopulations that produce more and the markers associated with that population means that they can be separated very easily,” Di Carlo said. “If we had very pure populations of cells that produced high levels of therapeutic proteins, we would have better treatments.”The nanovials are commercially available from Partillion Bioscience, a company co-founded by Di Carlo and founded in CNSI’s on-campus incubator. Expand.
Reference: “Correlating growth factor secretion in nanovials with single cell transcriptome using SEC-seq” Shreya Udani, Justin Langerman, Doyeon Koo, Sevana Baghdasarian, Brian Cheng, Simran Kang, Citradewi Soemardy, Joseph de Rutte, Kathrin Plath, Dino Di Carlo, December 11, 2023; natural nanotechnology.
DOI: 10.1038/s41565-023-01560-7The study’s lead author is Shreya Udani, who received her PhD from UCLA in 2023. Other co-authors, all at UCLA, are staff scientist Justin Langerman; Doyoung Koo, who received his Ph.D. in 2023. graduate students Sevana Bagdasarian and Chitradewi Somardi; undergraduate student Brian Chen; Simran Kang received her bachelor’s degree in 2023. and Joseph de Rutte, who completed his PhD in 2020 and is co-founder and CEO of Partillion.This research was supported by: National Institutes of Health It also won the Stem Cell Nanomedicine Program Award, jointly funded by CNSI and the Broad Center for Stem Cell Research.
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