In a collaborative study, researchers from Kyushu University and Harvard Medical School have identified proteins that can turn or “reprogram” fibroblasts — the most commonly found cells in skin and connective tissue — into cells with similar properties to limb progenitor cells. Publishing in Developmental Cell (“Direct reprogramming of non-limb fibroblasts to cells with properties of limb progenitors”), the researchers’ findings have enhanced our understanding of limb development and have set the stage for regenerative therapy in the future.
Globally, close to 60 million people are living with limb loss. Amputations can result from various medical conditions such as tumors, infections, and birth defects, or due to trauma from industrial accidents, traffic accidents, and natural disasters such as earthquakes. People with limb injuries often rely on synthetic materials and metal prostheses, but many researchers are studying the process of limb development, with the aim of bringing regenerative therapy, or natural tissue replacement, one step closer as a potential treatment.
“During limb development in the embryo, limb progenitor cells in the limb bud give rise to most of the different limb tissues, such as bone, muscle, cartilage and tendon. It’s therefore important to establish an easy and accessible way of making these cells,” explains Dr. Yuji Atsuta, lead researcher who began tackling this project at Harvard Medical School and continues it as a lecturer at Kyushu University’s Graduate School of Sciences.
Currently, a common way to obtain limb progenitor cells is directly from embryos, which, in the case of human embryos, raises ethical concerns. Alternatively, they can be made using induced pluripotent stem cells — adult cells which are reprogrammed into an embryonic-like state, and which can later be coaxed into specific tissue types. The new method developed by Atsuta and colleagues, which directly reprograms fibroblast cells into limb progenitor cells and bypasses induced pluripotent stem cells, simplifies the process and reduces costs. It also mitigates the concern of cells turning cancerous, which often occurs with induced pluripotent stem cells.
In the initial phase of the study, the researchers looked at what genes were expressed in the early limb buds in mice and chicken embryos. Almost all cells in the body, including fibroblasts and limb progenitor cells, contain identical genomic DNA, but the different properties and functions of each cell type emerge during development due to changes in gene expression (in other words, which genes are active, and which proteins are produced by the cell). One way that gene expression is controlled in cells is by specific proteins, called transcription factors.
The research group identified 18 genes, mostly transcription factors, that are more highly expressed in the early limb bud compared to other tissues. They cultured fibroblasts from mouse embryos and introduced these 18 genes into the fibroblasts using viral vectors so that the cells produced these 18 protein factors. They found that the modified fibroblasts took on the properties and showed similar gene expression to naturally-occurring limb progenitor cells found in limb buds.
Next, over a series of experiments, the researchers narrowed down their selection and determined that only three protein factors were essential to reprogram mouse fibroblasts into limb progenitor-like cells: Prdm16, Zbtb16, and Lin28a. A fourth protein, Lin41, helped the cultured limb progenitor cells grow and multiply more rapidly.
The researchers not only confirmed that the reprogrammed limb progenitor cells had similar gene expression to natural limb progenitor cells, but also had similar ability. “These reprogrammed cells are not only molecular mimics; we have confirmed their potential to develop into specialized limb tissues, both in laboratory dishes (in vitro) and also in living organisms (in vivo),” says Atsuta. “Testing in vivo was particularly challenging, as we had to transplant the reprogrammed mouse cells into the limb buds of chicken embryos.”
In these experiments, the researchers used lentiviruses, which insert genes directly into the infected cells’ genome, raising the risk that the cells can become cancer. Instead, the team is considering other safer vectors, such as adeno-associated viruses or plasmids, which deliver genes to the cells without inserting genes into the genome.
Atsuta’s lab group is now trying to apply this method to human cells, for future therapeutic applications, and also to snakes, whose ancestors had limbs that were subsequently lost during evolution. “Interestingly, the reprogrammed limb progenitor cells generated limb bud-like organoids, so it seems possible to generate limb tissues in species that no longer possess them. The study of limbless snakes can uncover new pathways and knowledge in developmental biology.”

News
Why ‘Peniaphobia’ Is Exploding Among Young People (And Why We Should Be Concerned)
An insidious illness is taking hold among a growing proportion of young people. Little known to the general public, peniaphobia—the fear of becoming poor—is gaining ground among teens and young adults. Discover the causes [...]
Team finds flawed data in recent study relevant to coronavirus antiviral development
The COVID pandemic illustrated how urgently we need antiviral medications capable of treating coronavirus infections. To aid this effort, researchers quickly homed in on part of SARS-CoV-2's molecular structure known as the NiRAN domain—an [...]
Drug-Coated Neural Implants Reduce Immune Rejection
Summary: A new study shows that coating neural prosthetic implants with the anti-inflammatory drug dexamethasone helps reduce the body’s immune response and scar tissue formation. This strategy enhances the long-term performance and stability of electrodes [...]
Scientists discover cancer-fighting bacteria that ‘soak up’ forever chemicals in the body
A family of healthy bacteria may help 'soak up' toxic forever chemicals in the body, warding off their cancerous effects. Forever chemicals, also known as PFAS (per- and polyfluoroalkyl substances), are toxic chemicals that [...]
Johns Hopkins Researchers Uncover a New Way To Kill Cancer Cells
A new study reveals that blocking ribosomal RNA production rewires cancer cell behavior and could help treat genetically unstable tumors. Researchers at the Johns Hopkins Kimmel Cancer Center and the Department of Radiation Oncology and Molecular [...]
AI matches doctors in mapping lung tumors for radiation therapy
In radiation therapy, precision can save lives. Oncologists must carefully map the size and location of a tumor before delivering high-dose radiation to destroy cancer cells while sparing healthy tissue. But this process, called [...]
Scientists Finally “See” Key Protein That Controls Inflammation
Researchers used advanced microscopy to uncover important protein structures. For the first time, two important protein structures in the human body are being visualized, thanks in part to cutting-edge technology at the University of [...]
AI tool detects 9 types of dementia from a single brain scan
Mayo Clinic researchers have developed a new artificial intelligence (AI) tool that helps clinicians identify brain activity patterns linked to nine types of dementia, including Alzheimer's disease, using a single, widely available scan—a transformative [...]
Is plastic packaging putting more than just food on your plate?
New research reveals that common food packaging and utensils can shed microscopic plastics into our food, prompting urgent calls for stricter testing and updated regulations to protect public health. Beyond microplastics: The analysis intentionally [...]
Aging Spreads Through the Bloodstream
Summary: New research reveals that aging isn’t just a local cellular process—it can spread throughout the body via the bloodstream. A redox-sensitive protein called ReHMGB1, secreted by senescent cells, was found to trigger aging features [...]
AI and nanomedicine find rare biomarkers for prostrate cancer and atherosclerosis
Imagine a stadium packed with 75,000 fans, all wearing green and white jerseys—except one person in a solid green shirt. Finding that person would be tough. That's how hard it is for scientists to [...]
Are Pesticides Breeding the Next Pandemic? Experts Warn of Fungal Superbugs
Fungicides used in agriculture have been linked to an increase in resistance to antifungal drugs in both humans and animals. Fungal infections are on the rise, and two UC Davis infectious disease experts, Dr. George Thompson [...]
Scientists Crack the 500-Million-Year-Old Code That Controls Your Immune System
A collaborative team from Penn Medicine and Penn Engineering has uncovered the mathematical principles behind a 500-million-year-old protein network that determines whether foreign materials are recognized as friend or foe. How does your body [...]
Team discovers how tiny parts of cells stay organized, new insights for blocking cancer growth
A team of international researchers led by scientists at City of Hope provides the most thorough account yet of an elusive target for cancer treatment. Published in Science Advances, the study suggests a complex signaling [...]
Nanomaterials in Ophthalmology: A Review
Eye diseases are becoming more common. In 2020, over 250 million people had mild vision problems, and 295 million experienced moderate to severe ocular conditions. In response, researchers are turning to nanotechnology and nanomaterials—tools that are transforming [...]
Natural Plant Extract Removes up to 90% of Microplastics From Water
Researchers found that natural polymers derived from okra and fenugreek are highly effective at removing microplastics from water. The same sticky substances that make okra slimy and give fenugreek its gel-like texture could help [...]