New Regulatory Mechanism Discovered for Cell Identity and Behavior in Forming Organs

Genes & Development study from Cincinnati Childrens May Have Broad Implications for Cancer, Other Diseases

Friday, October 31, 2008

Two proteins interact in a previously unknown molecular mechanism that may have broad implications in future studies looking at the causes of defective organs in fetuses, metastatic cancers and other diseases, according to researchers at Cincinnati Childrens Hospital Medical Center.
Reporting their work in the Nov. 1 Genes & Development, the researchers said the mechanism coordinates cell identity and behavior in the forming organs of embryos.

Our study helps address the current challenge of finding out where cell specificity comes from, how cells do what they do in the context of disease and development, and how these activities are regulated, said Aaron Zorn, PhD, a researcher in the Division of Developmental Biology at Cincinnati Childrens and the study's corresponding author. This helps inform research into how we tell early stem cells what to become. If someone has diabetes, for example, how do we tell a cell to become a pancreas cell so it will produce insulin?

The study involved embryos of Xenopus frogs, a species indigenous to Africa often used in early biomedical studies. The scientists discovered a signaling protein very common in developmental biology, Wnt11 (Wingless), has to be inhibited by the modulating protein Sfrp5 (Secreted Frizzled Related Protein), a known antagonist of Wnt. Without this restriction, Wnt signaling runs amok and the frogs foregut, liver and pancreas form improperly from a cascade of disorganized cell growth.

We point out that Wnt has two key roles here - one controlling the cell expression pathway to tell cells what they are supposed to be, and the other controlling the pathway for cell movement, behavior and adhesion, said Dr. Zorn, also associate professor of pediatrics at the University of Cincinnati (UC) College of Medicine. Without Sfrp5 controlling what Wnt does in both pathways, things go horribly wrong in the developing foregut and its organs.

The Wnt signaling pathway is a complex network of proteins best known for their role in stimulating cell behavior during embryo development and in cancer. They also are involved in normal physiological processes in adult animals. Parts of the Wnt pathway have been conserved between species during the long course of evolution, all the way from simple roundworms to humans.

Previous research in Xenopus has established that a low level of activity from a molecule called B-catenin - which promotes cell-to-cell adhesion and is part of the Wnt pathway - is necessary to maintain accurate foregut formation and initiate liver and pancreas development. Unknown before the study by Dr. Zorns team was which Wnt genes are involved and how Wnt and B-catenin activity are regulated along the frogs developing anterior-posterior body axis.

During the very early phases of embryo development - when the organism is still essentially flattened layers of cells called an endoderm - Dr. Zorns team found Wnts stimulation of B-catenin must be restricted in the anterior region so the tissue of forming foregut organs maintain its integrity. Their experiments showed that Sfrp5 steps in at the right time and place to repress Wnt signaling, allowing the cells to form an epithelial sheet, or lining - an essential step in organ development.

In one experiment, when researchers removed the Sfrp5 protein, the resulting Sfrp5-depleted Xenopus embryos had smaller foregut cavities filled with unorganized early-stage endoderm cells, which were incapable of properly forming liver and pancreatic organs.

Dr. Zorn and colleagues said their results have possible implications in metastatic cancer. For one, Sfrp proteins are already known to be tumor suppressors that are genetically inactivated in some cancers as they progress to aggressive carcinomas. Carcinomas typically originate in epithelial cells - which form linings surrounding the surfaces and cavities of many body structures - then spread into surrounding organs and tissues.

In cancer development, the research team is suggesting a loss of Sfrp function may unleash Wnt to trigger elevated B-catenin expression, allowing its stimulation of cell-to-cell adhesion to proliferate quickly. Rapid cell proliferation and adhesion are common in cancerous and pre-cancerous conditions. It could also let Wnt send improper signals that cause a loss of structural integrity in epithelial cells, allowing cancer to spread, or metastasize.

We talked about this mechanism in the context of cancer because the control of cell specificity, and of movement and behavior, also occurs in cancer, Dr. Zorn said. Cells will start to proliferate out of control and then, when a cancer starts to go metastatic, they will also start to change behavior. They become motile, moving spontaneously and actively, and they become invasive.

The early stage nature of the study means it would be premature to suggest the Wnt11-Sfrp5 mechanism might become the basis of diagnostic or therapeutic strategies for patients, Dr. Zorn said. The next step is to use these results as a basis for future studies, probably involving mice, to verify the mechanisms applicability to mammalian embryo development and see how it affects disease, he said.

Participating in the study were the Cincinnati Childrens Research Foundation; Department of Pediatrics, UC College of Medicine; Department of Cell Biology and Anatomy, University of Arizona Health Sciences Center and the State Key Laboratory of Phytochemistry and Plant Resources at the Kunming Institute of Botany, Kunming, China. Other researchers include lead author, Yan Li, and Scott A. Rankin, Debora Sinner, Alan P. Kenney and Paul A. Kreig.

Funding support came from the National Institutes of Health.

Cincinnati Children's Hospital Medical Center is one of Americas top three children's hospitals for general pediatrics and is highly ranked for its expertise in digestive diseases, respiratory diseases, cancer, neonatal care, heart care and neurosurgery, according to the annual ranking of best children's hospitals by U.S. News & World Report. One of the three largest children's hospitals in the U.S., Cincinnati Childrens is affiliated with the University of Cincinnati College of Medicine and is one of the top two recipients of pediatric research grants from the National Institutes of Health. For its achievements in transforming healthcare, Cincinnati Children's is one of six U.S. hospitals since 2002 to be awarded the American Hospital Association-McKesson Quest for Quality Prize ® for leadership and innovation in quality, safety and commitment to patient care. The hospital is a national and international referral center for complex cases, so that children with the most difficult-to-treat diseases and conditions receive the most advanced care leading to better outcomes.