What are the key genes of Wnt secretion?
Understanding of cancer initiation and development is one of the biggest tasks we human are facing. Tumorigenesis is a systematically functional alteration on individual.
Wnt signaling pathway regulates cell proliferation and induces morphological changes. The downstream effect of Wnt ligands is known to be highly activated in tumor. Its alteration occurs in almost all the cancer types, especially in colorectal cancer. But, unfortunately, we know very little about Wnt secretion process, the upstream of Wnt signaling pathway. It requires more attention and efforts for finding some key genes.
Combinations of microfluidics and synthetic biology inspire and excite us. We try to design and apply new methods to study cell-cell interactions in foundational researches and also build up high-throughput microfluidic methods for synthetic biology.
In a word
In this project, we aim to track the Wnt secretion pathway using microfluidic Donor-Reporter system via synthetic biology approaches to reveal some potential driver genes that lead to the initiation of tumor and some downstream effects in Wnt-secretion during tumorigenesis. At the same time, we build a predictive model for Wnt secretion and signaling network.
Microfluidic Donor-Reporter system
To study Wnt secretion, we need to develop a microfluidic based Donor-Reporter system, which including construction of Wnt reporter cell line and Wnt secreting cell line via synthetic biology approaches. Using CRISPR-Cas9 system to knock out genes in Wnt secreting cells, sequential response on reporter cells can show the Wnt secretion capability via fluorescence to reveal the genetic effects of the knockout gene. FACS will be used to sort out the potential gene that play essential roles in Wnt secretion.
Here two new microfluidic methods are now developing for high-throughput screening of Wnt secretion related genes using microfluidics. One is double emulsion system and the other is micro-well system. Those two systems have their benefits and in the future, we want to apply it to not only study of one specific signaling pathway but also cell-cell interactions. The newly designed techniques can also be applied to build up high-throughput methods for synthetic biology.
Figure 1 Wnt signaling pathway
Figure 2 Microfluidic chip
Figure 3 Chip graph(Left) & Double emulsion droplets generation(Right)
Prediction and network modeling
At the same time, we want to build a model that associated with the origin and the effects of Wnt protein in tumor's environment. We try to find key genes associated with Wnt secretion by using gene co-expression network analysis. Then we explore the alteration of those genes and their potential function by comparative genomics approach. Here, we use data from TCGA(The Cancer Genome Atlas) for Wnt signaling network analysis.
Figure 4 Simplified model of Wnt secretion and its downstream effects
Figure 1 comes from the Wnt Homepage of Roel Nusse Lab.
Figure 2 and Figure 3 come from Dolomite corporation.