Genetically Modified Cell Therapy for the Reduction of Lung Injury
Lead Research Organisation:
University College London
Department Name: Medicine
Abstract
Chronic lung diseases are extremely common. Many lung diseases cause destruction of the gas exchange surface of the lung (the lung parenchyma) making patients progressively breathless until they reach respiratory failure and die. Current medical interventions are failing these patients, leading to a search for new treatments.
It appears that the lung not only uses resident cells to affect repair after damage, but also recruits circulating cells from the bone marrow. We believe these cells may be used to deliver genetic therapies into the injured lung parenchyma. We have two major goals:
1. To determine which bone marrow cells are responsible for lung repair.
2. To use gene therapy to make our chosen population of bone marrow cells produce a growth factor, enhancing lung repair.
We believe the combined expertise of scientists and clinicians at the Centre for Respiratory Disease (UCL) and the Cellular and Gene therapy Unit (Institute of Child Health, UCL) will lead to novel treatments for a wide range of debilitating diseases including emphysema and lung fibrosis.
It appears that the lung not only uses resident cells to affect repair after damage, but also recruits circulating cells from the bone marrow. We believe these cells may be used to deliver genetic therapies into the injured lung parenchyma. We have two major goals:
1. To determine which bone marrow cells are responsible for lung repair.
2. To use gene therapy to make our chosen population of bone marrow cells produce a growth factor, enhancing lung repair.
We believe the combined expertise of scientists and clinicians at the Centre for Respiratory Disease (UCL) and the Cellular and Gene therapy Unit (Institute of Child Health, UCL) will lead to novel treatments for a wide range of debilitating diseases including emphysema and lung fibrosis.
Technical Summary
Many common diseases of the gas exchange surface of the lung (the parenchyma) have no specific treatment but result in significant morbidity and mortality. Idiopathic Pulmonary Fibrosis (IPF) is characterized by alveolar epithelial cell injury, interstitial inflammation, fibroblast proliferation and collagen accumulation within the lung parenchyma. It has been recently discovered that post injury the lung not only uses resident cells but also recruits circulating bone marrow-derived cells (BMDC). However the source of these cells and the mechanism by which repair occurs is unknown. Keratinocyte Growth Factor (KGF) is a critical factor stimulating epithelial proliferation during lung repair. This proposal aims to examine the role of BMDCs in repairing acute parenchymal lung injury. We hypothesise that targeted therapy for lung injury could be performed based on the recruitment of a modified population of BMDCs that promote epithelial repair. We have three key objectives:
1. To determine the efficiency and time course of conditional lentiviral KGF expression in different BMDC populations.
2. To determine which sub-populations of BMDCs transduced with a conditional lentivirus expressing KGF increase type II pneumocyte proliferation in vitro and in vivo.
3. To determine whether the intra-tracheal or systemic delivery of bone marrow cell sub-populations over-expressing KGF can improve lung repair after injury.
In preliminary data we show that haematopoietic stem cell transplants can deliver KGF to injured lung parenchyma resulting in type II pneumocyte proliferation and attenuation of bleomycin induced lung fibrosis. I wish to further characterise this effect and determine whether a sub-population of daughter cells could be used to the same effect without the necessity for bone marrow transplantation enabling a clinically useful therapy.
We will use established flow cytometry and cell culture techniques to culture haematopoietic stem cells, mesenchymal stem cells and macrophages and then transduce them with a tetracycline controlled lentivirus expressing KGF. I will then use transduced cells activated with doxycycline in vitro to examine type II pneumocyte mitogenic response. The cell sub-populations will then be systemically delivered, or in the case of macrophages both systemically and intra-tracheally, both with and without bleomycin-induced lung damage. Effects on type II cell proliferation and attenuation of lung damage will be assessed by lung morphology, inflammation and collagen load (hydroxyproline content).
We believe our proposal may lead to a completely novel cell based therapy and a new means of delivering gene therapy to the lung, improving the treatment of currently intractable diseases.
1. To determine the efficiency and time course of conditional lentiviral KGF expression in different BMDC populations.
2. To determine which sub-populations of BMDCs transduced with a conditional lentivirus expressing KGF increase type II pneumocyte proliferation in vitro and in vivo.
3. To determine whether the intra-tracheal or systemic delivery of bone marrow cell sub-populations over-expressing KGF can improve lung repair after injury.
In preliminary data we show that haematopoietic stem cell transplants can deliver KGF to injured lung parenchyma resulting in type II pneumocyte proliferation and attenuation of bleomycin induced lung fibrosis. I wish to further characterise this effect and determine whether a sub-population of daughter cells could be used to the same effect without the necessity for bone marrow transplantation enabling a clinically useful therapy.
We will use established flow cytometry and cell culture techniques to culture haematopoietic stem cells, mesenchymal stem cells and macrophages and then transduce them with a tetracycline controlled lentivirus expressing KGF. I will then use transduced cells activated with doxycycline in vitro to examine type II pneumocyte mitogenic response. The cell sub-populations will then be systemically delivered, or in the case of macrophages both systemically and intra-tracheally, both with and without bleomycin-induced lung damage. Effects on type II cell proliferation and attenuation of lung damage will be assessed by lung morphology, inflammation and collagen load (hydroxyproline content).
We believe our proposal may lead to a completely novel cell based therapy and a new means of delivering gene therapy to the lung, improving the treatment of currently intractable diseases.