Team:Purdue/Project

From 2009.igem.org

(Difference between revisions)
(The Concept)
Line 29: Line 29:
=== The Concept ===
=== The Concept ===
-
The basic idea is to use the naturally mobility of a microglial cell to find a specific cell found in 1% of some glioblastoma multiforme.
+
Our basic goal is to use the natural mobility of a microglial cell to find specific cells found in 1% of some glioblastoma multiforme.
 +
Glioblastoma multiforme (GBM) is a deadly brain cancer found characterizing over 60% of brain cancers.  With current treatment options, even the best-cared for patient, once diagnosed, has little over one year to live.  Most recently, Ted Kennedy died of this cancer.
-
Microglial cells are the macrophage of the central nervous system. They are needed due to the blood brain barrier that is necessary for our system to work. In a resting state, the microglia have extensions that are constantly surveying the environment. The microglia interpret signals from the environment and act accordingly. The microglia have the ability to "clean up" dead cells and debris via endocytosis.  
+
Scientists have found that GBM is made up of many different cell types, some of which make this primary tumor difficult to destroy. In the center of some of these tumor masses, there is a small subset of cells that display stem cell-like qualities. These cells express CD133, a protein found only on stem cells. These stem-like cells have been shown to be resistant to hypoxia, irradiation, and some chemotherapies.  These qualities indicate to some a possible reason why recurrence is so high in patients diagnosed with GBM: it is possible that these stem-like cells are the crucial tumor-initiating cells (Cancer Stem Cell Theory), but they are not destroyed along with rest of the tumor mass using current therapies.
-
Glioblastoma multiforme is made up of many different cell types that make this primary tumor difficult to destroy. Also, in the center of some of these tumor masses, scientists have found a small subset of cells that display stem cell-like qualities. They have been shown to be resistant to hypoxia, irradiation, and some chemo-therapies which indicate to some as a possible reason why recurrence is so high in patients diagnosed with glioblastoma multiforme.
+
Microglial cells are the macrophage of the central nervous system. In a resting state, the microglia have extensions that constantly survey the environment. The microglia interpret signals from the environment and act accordingly. Also, microglia have the ability to "clean up" dead cells and debris via phagocytosis, and move around towards possible threats.  
-
We plan to achieve this goal by creating a chimeric receptor that is specific for the 1% subset of cells we are attacking. This small population is CD133 positive. This protein is a marker found on some stem cells, but as of right now no one has fully characterized CD133. We will use part of the CD133 antibody (Fab region) and most of the Fc gamma Receptor to create this very specific chimeric receptor. The Fc Receptor is native to the microglia, and thus have the pathway needed to reach the nucleus. As of right now, we are planning on using the C-Jun promoter to create our tat-GFP fusion protein.  
+
Our goal is to use the microglia to seek out the CD133+ stem-like cells found in the glioblastoma.  We plan to achieve this goal by creating a chimeric receptor that is specific for the CD133 protein. Currently, the CD133 protein is not fully characterized. To solve this dilemma, we will use part of the CD133 antibody (Fab region) and most of the Fc gamma receptor to create a very specific chimeric receptor. The Fc receptor is native to the microglia, and thus the microglia have the pathway needed to activate a signal cascade, of which a downstream product is the protein c-jun. As of right now, we are planning on using the c-jun promoter to activate our product, a tat-GFP fusion protein.
-
Tat is a cell penetrating peptide that was first discovered by two independent labs studying HIV-1. While the mechanism is not quite known, tat has the ability to cross cell membranes with ease. Using this inherent quality we want to attach it to GFP to create a fusion protein that can label the CD133 cells. The idea being, if it can be labeled it can be killed.  
+
Tat is a cell-penetrating peptide that was first discovered by two independent labs studying HIV-1. Although the mechanism is unknown, tat has the ability to cross cell membranes with ease. Using this inherent quality, we are attaching it to GFP to create a fusion protein that would label the CD133 cells. The basic idea is that if the CD133+ cell can be labeled, it can be killed.  
-
In the future, we would hook up an apoptitic factor to the tat to attempt to kill the cells.  
+
In the future, we would like to hook up an apoptotic factor to the tat in an attempt to kill the CD133+ glioblastoma cells (as opposed to just labeling them).  
Overall, we have created a seek and destroy system that is modular and can be adapted to other problems in the human body.
Overall, we have created a seek and destroy system that is modular and can be adapted to other problems in the human body.

Revision as of 17:20, 20 October 2009

Header.png

Home The Team The Project Parts Submitted to the Registry Modeling Notebook Safety


Contents

Abstract

Glioblastoma multiforme (GBM) is one of the most common forms of primary brain cancer, which usually results in fatality. To date, it has been difficult to overcome primary brain cancer resulting from GBM, primarily because the cancer-initiating cells are suspected to be highly resistant to current cancer therapies. The present study is focused on CD133+ cells found in primary GBM samples. CD133+ cells have shown resistance to hypoxia, irradiation, and some forms of chemotherapy. CD133+ hunting machines will be created by genetically engineering microglial cells (BV-2) with two constructs using mammalian expression vectors. The project will also take advantage of inherent qualities of the microglia such as constant environmental sensing and quick motility. The engineered BV-2s will be equipped to locate the specific GBMs and label the targeted cells with a tat-GFP fusion protein. To create an in-vivo-like environment, the cells will be grown in 3D collagen media. This would create a maze and challenge the microglia to seek out the cancer cells. Cell sorting techniques will be used to measure the accuracy in hitting positive targets, the CD133+ cells. It is the goal of this study to show an alternative approach to cancer treatment, and to emphasize the power of biologically available options to fight the disease.

Project Details

JessSciencePU.JPG
Jessamine's doing science!



The Concept

Our basic goal is to use the natural mobility of a microglial cell to find specific cells found in 1% of some glioblastoma multiforme.

Glioblastoma multiforme (GBM) is a deadly brain cancer found characterizing over 60% of brain cancers. With current treatment options, even the best-cared for patient, once diagnosed, has little over one year to live. Most recently, Ted Kennedy died of this cancer.

Scientists have found that GBM is made up of many different cell types, some of which make this primary tumor difficult to destroy. In the center of some of these tumor masses, there is a small subset of cells that display stem cell-like qualities. These cells express CD133, a protein found only on stem cells. These stem-like cells have been shown to be resistant to hypoxia, irradiation, and some chemotherapies. These qualities indicate to some a possible reason why recurrence is so high in patients diagnosed with GBM: it is possible that these stem-like cells are the crucial tumor-initiating cells (Cancer Stem Cell Theory), but they are not destroyed along with rest of the tumor mass using current therapies.

Microglial cells are the macrophage of the central nervous system. In a resting state, the microglia have extensions that constantly survey the environment. The microglia interpret signals from the environment and act accordingly. Also, microglia have the ability to "clean up" dead cells and debris via phagocytosis, and move around towards possible threats.

Our goal is to use the microglia to seek out the CD133+ stem-like cells found in the glioblastoma. We plan to achieve this goal by creating a chimeric receptor that is specific for the CD133 protein. Currently, the CD133 protein is not fully characterized. To solve this dilemma, we will use part of the CD133 antibody (Fab region) and most of the Fc gamma receptor to create a very specific chimeric receptor. The Fc receptor is native to the microglia, and thus the microglia have the pathway needed to activate a signal cascade, of which a downstream product is the protein c-jun. As of right now, we are planning on using the c-jun promoter to activate our product, a tat-GFP fusion protein.

Tat is a cell-penetrating peptide that was first discovered by two independent labs studying HIV-1. Although the mechanism is unknown, tat has the ability to cross cell membranes with ease. Using this inherent quality, we are attaching it to GFP to create a fusion protein that would label the CD133 cells. The basic idea is that if the CD133+ cell can be labeled, it can be killed.

In the future, we would like to hook up an apoptotic factor to the tat in an attempt to kill the CD133+ glioblastoma cells (as opposed to just labeling them).

Overall, we have created a seek and destroy system that is modular and can be adapted to other problems in the human body.

Plasmid Design

What We've Done

Future Plans

References