Authors: Adam Wanner; Laurent P. Nicod; Andre Perruchoud; William MacNee

Author: Vladimir R. Muzykantov

Authors: Robin S. Dothager; David Piwnica-Worms

Characterization and noninvasive measurement of molecular pathways and biochemistry in living cells, animal models, and humans at the cellular and molecular level is now possible using remote imaging detectors. Positron and single photon emission tomography scanners, highly sensitive cameras for bioluminescence and fluorescence imaging, as well as high-magnetic-field magnetic resonance imaging scanners, can be used to study such diverse processes as signal transduction, receptor density and function, host response to pathogens, cell trafficking, and gene transfer. In many cases, images from more than one modality can be fused, allowing structure–function and multifunction relationships to be studied on a tissue-restricted or regional basis. “Molecular imaging” holds enormous potential for elucidating the molecular mechanisms of pulmonary disease and therapeutic response in intact animal models and humans.

Authors: Ricardo J. Giordano; Julianna K. Edwards; Rubin M. Tuder; Wadih Arap; Renata Pasqualini

Phage display of random peptide libraries is a powerful, unbiased method frequently used to discover ligands for virtually any protein of interest and to reveal functional protein–protein interaction partners. Moreover, in vivo phage display permits selection of peptides that bind specifically to different vascular beds without any previous knowledge pertaining to the nature of their corresponding receptors. Vascular targeting exploits molecular differences inherent in blood vessels within given organs and tissues, as well as diversity between normal and angiogenic blood vessels. Over the years, our group has identified phage capable of homing to lung blood vessels based on screenings using immortalized lung endothelial cells combined with in vivo selections after intravenous administration of combinatorial libraries. Peptides targeting lung vasculature have been extensively characterized and a lead homing peptide has shown interesting biological properties, bringing novel insights as to the implications of lung endothelial cell apoptosis in the pathogenesis of emphysema. We have also designed and developed targeted nanoparticles with imaging capabilities and/or drug delivery functions by combining phage display technology and elemental gold (Au) nanoparticles, constituting a promising platform for the development of therapeutic agents for imaging and treatment of lung disorders. Given the important role of the endothelium in the pathogenesis and progression of several diseases associated with the airways, ligand-directed discovery of lung vascular markers is an important milestone toward the development of future targeted therapies.

Author: Vasilis Ntziachristos

Biomedical imaging has become an important tool in the study of “-omics” fields by allowing the noninvasive visualization of functional and molecular events using in vivo staining and reporter gene approaches. This capacity can go beyond the understanding of the genetic basis and phenotype of such respiratory conditions as acute bronchitis, adult respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease (COPD), and asthma and investigate the development of disease and of therapeutic events longitudinally and in unperturbed environments. Herein, we show how the application of novel quantitative optical imaging methods, using transillumination and fluorescence molecular tomography (FMT), can allow visualization of pulmonary inflammation in small animals in vivo. The results confirm prior observations using a protease-sensitive probe. We discuss how this approach enables in vivo insights at the system level as to the dynamic role of proteases in respiratory pathophysiology and their potential as therapeutic targets. Overall, the proposed imaging method can be used with a significantly wider range of possible targets and applications in lung imaging.

Authors: Kerri A. Massey; Jan E. Schnitzer

A major goal of molecular medicine is to target imaging agents or therapeutic compounds to a single organ. Targeting imaging agents to a single organ could facilitate the high-resolution, in vivo imaging of molecular events. In addition, genetic and acquired diseases primary to a single organ, such as cystic fibrosis, tuberculosis, lung cancer, pulmonary fibrosis, pulmonary hypertension, and acute respiratory distress syndrome, could be specifically targeted in the lung. By targeting and concentrating imaging agents or therapeutics to the lungs, deleterious side effects can be avoided with greater efficacy at much lower dosages. Pathologic changes can be identified earlier and followed over time. In addition, therapeutics that have been abandoned due to toxicities may find renewed utility when coupled with specific targeting agents such as antibodies. To achieve these goals, distinct molecular signatures must be found for each organ or disease-state.

Authors: Kiarash Emami; Michael Stephen; Stephen Kadlecek; Robert V. Cadman; Masaru Ishii; Rahim R. Rizi

Improvements in the quantitative assessment of structure, function, and metabolic activity in the lung, combined with improvements in the spatial resolution of those assessments, enhance the diagnosis and evaluation of pulmonary disorders. Radiologic methods are among the most attractive techniques for the comprehensive assessment of the lung, as they allow quantitative assessment of this organ through measurements of a number of structural, functional, and metabolic parameters. Hyperpolarized nuclei magnetic resonance imaging (MRI) has opened up new territories for the quantitative assessment of lung function and structure with an unprecedented spatial resolution and sensitivity. This review article presents a survey of recent developments in the field of pulmonary imaging using hyperpolarized nuclei MRI for quantitative imaging of different aspects of the lung, as well as preclinical applications of these techniques to diagnose and evaluate specific pulmonary diseases. After presenting a brief overview of various hyperpolarization techniques, this survey divides the research activities of the field into four broad areas: lung microstructure, ventilation, oxygenation, and perfusion. Finally, it discusses the challenges currently faced by researchers in this field to translate this rich body of methodology into wider-scale clinical applications.

Authors: Harvey O. Coxson; Stephen Lam

Ever since the site and nature of airflow obstruction in chronic obstructive pulmonary disease was described by Hogg, Thurlbeck, and Macklem, investigators have been looking for methods to noninvasively measure the airway wall dimensions. Recent advances in computed tomography technology and new computer algorithms have made it possible to visualize and measure the airway wall and lumen without the need for tissue. However, while there is great hope for computed tomographic assessment of airways, it is well known that the spatial resolution does not allow small airways to be visualized and there are still concerns about the sensitivity of these measurements obtained from these airways. Optical coherence tomography is a new bronchoscopic imaging technique that has generated considerable interest because the spatial resolution is much higher than computed tomography. While relatively more invasive than computed tomography, it has the advantage of not exposing the patient to ionizing radiation. This review discusses some of the data surrounding these two imaging techniques in patients with chronic obstructive pulmonary disease. These imaging techniques are extremely important in the assessment of patients with chronic obstructive pulmonary disease because therapy that is designed to modulate the inflammation in airways may be contraindicated in subjects with the emphysema phenotype and visa versa. Therefore, these new imaging techniques are very likely to play a front-line role in the study of chronic obstructive pulmonary disease and will, hopefully, allow clinicians to phenotype individuals, thereby personalizing their treatment.

Authors: Luc Thiberville; Mathieu Salaün; Samy Lachkar; Stephane Dominique; Sophie Moreno-Swirc; Christine Vever-Bizet; Genevieve Bourg-Heckly

Confocal endomicroscopes aim at providing to the clinician microscopic imaging of a living tissue. The currently available microendoscopic devices use the principle of confocal fluorescent microscopy, in which the objective is replaced by an optical fiber and a miniaturized scanhead at the distal end of the endoscope or by a retractable bundle of optical fibers. Such systems have recently been applied to the explorations of several organs, including the gastrointestinal tract, and more recently to the proximal and distal airways in vivo. Respiratory fluorescence microendoscopes use 488 nm or 660 nm excitation laser light and thin flexible miniprobes that are introduced into the working channel of the bronchoscope. The devices have a lateral resolution of 3 μm, a field of view of 600 μm, and produce real-time imaging at 9 frames per second. For in vivo imaging, the miniprobe is applied onto the bronchial wall surface or advanced into a distal bronchiole down to the acinus. In nonsmokers, the 488-nm excitation device images the autofluorescence of the elastin that is contained in the basement membrane of the proximal airways and that participates to the axial backbone of the peripheral interstitial respiratory system. In smokers, a specific tobacco tar–induced fluorescence allows in vivo macrophage and alveolar wall imaging. Using 660 nm excitation and topical methylene blue, the technique enables cellular imaging of both bronchial epithelial layer and peripheral lung nodules. This article reviews the capabilities and possible limitations of confocal microendoscopy for in vivo proximal and distal lung explorations.

Authors: Ke Zhang; Huafeng Fang; Gang Shen; John-Stephen A. Taylor; Karen L. Wooley

This mini-review highlights developments that have been made over the past year to advance the construction of well-defined nanoscale objects to serve as devices for cell transfection. Design of the nanoscale objects originated from biomimicry concepts, using histones as the model, to afford cationic shell crosslinked knedel-like (cSCK) nanoparticles. Packaging and delivery of plasmid DNA, oligonucleotides, and peptide nucleic acids were studied by dynamic light scattering, transmission electron microscopy, gel electrophoresis, biological activity assays, RT-PCR measurements, flow cytometry, and confocal fluorescence microscopy. With the demonstration of more efficient cell transfection in vitro than that achieved using commercially-available transfection agents, together with the other features offered by the robust nanostructural framework, work continues toward the application of these cSCKs for in vivo molecular recognition of genetic material, for imaging and therapy targeted specifically to pulmonary injury and disease.

Authors: Hans-Ulrich Kauczor; Julia Ley-Zaporozhan; Sebastian Ley

Magnetic resonance imaging (MRI) of the lung has shown tremendous progress in recent years. This includes parallel imaging, new contrast agents and mechanisms, ultrafast imaging, and respiratory gating. With these improvements in speed and image quality, MRI is now ready for routine clinical use. The main advantage for MRI of the lung is its unique combination of structural and functional assessment within a single imaging examination. This comprehensive imaging assessment is an asset when compared with computed tomography, which is complemented by the fact that MRI does not carry any exposure to ionizing radiation, making it especially advantageous in children, young adults, and for follow-up examinations either in disease surveillance or therapy monitoring. Clinical indications for MRI are: pulmonary vascular disease, especially pulmonary hypertension, airway diseases, especially cystic fibrosis; neoplastic disease, including staging of lung cancer as an alternative imaging modality; all pediatric indications (e.g., congenital anomalies); as well as follow-up examinations. Under investigation is the application of MRI for chronic obstructive pulmonary disease as well as asthma. In this regard the additional benefit from MRI using hyperpolarized gases has to be determined.

Author: Chaitanya R. Divgi

Molecular imaging (MI) may be defined as imaging in vivo using molecules that report on biologic function. This review will focus on the clinical use of radioactive tracers (nonpharmacologic amounts of compounds labeled with a radioactive substance) that permit external imaging using single photon emission computed tomography (planar, SPECT) or positron emission tomography (PET) imaging. Imaging of lung cancer has been revolutionized with the use of fluorine-18–labeled fluorodeoxyglucose (¹⁸F-FDG), an analog of glucose that can be imaged using PET. The ability to carry out whole body imaging after intravenous injection of 18F-FDG allows accurate staging of disease, helping to determine regional and distant nodal and other parenchymal involvement. Glycolysis is increased in nonmalignant conditions, including inflammation (e.g., sarcoidosis), and ¹⁸F-FDG PET is a sensitive method for evaluation of active inflammatory disease. Inflammatory disease has been imaged, even before the advent of PET, with planar and SPECT imaging using gallium-67, a radiometal that binds to transferrin. Metabolic alteration in pulmonary pathology is currently being studied, largely in lung cancer, primarily with PET, with a variety of other radiotracers. Prominent among these is thymidine; fluorine-18–labeled thymidine PET is being increasingly used to evaluate proliferation rate in lung and other cancers. This overview will focus on the clinical utility of ¹⁸F-FDG PET in the staging and therapy evaluation of lung cancer as well as in imaging of nonmalignant pulmonary conditions. PET and SPECT imaging with other radiotracers of interest will also be reviewed. Future directions in PET imaging of pulmonary pathophysiology will also be explored.

Author: Francis G. Blankenberg

Annexin V is a ubiquitous intracellular protein in humans that has a variety of intriguing characteristics, including a nanomolar affinity for the membrane-bound constitutive anionic phospholipid known as phosphatidylserine (PS). PS is selectively expressed on the surface of apoptotic or physiologically stressed cells. As such, radiolabeled forms of annexin V have been used in both animal models and human Phase I and Phase II trials to determine if this tracer can be employed as an early surrogate marker of therapeutic efficacy in NSCLC and non-Hodgkin's lymphoma. Many other pulmonary imaging applications of radiolabeled annexin V are also possible, including the detection and monitoring of active pulmonary inflammation and other pathophysiologic stressors in a variety of diseases. In this article, the salient molecular features of apoptosis (and other forms of cell death) that permits imaging with radiolabeled annexin V will be discussed. The latest results from Phase II imaging trials with NSCLC and non-Hodgkin's lymphoma will be also be detailed. Finally, the potential future application of this tracer for the imaging of other pulmonary pathologies will be outlined.

Author: Myrna B. Dolovich

¹⁸F-FDG positron emission tomographic (PET) scanning is a major imaging tool widely used to investigate lung function and lung disease. Tomographic imaging of drug delivered to the lung via the aerosol route can provide data that link the regional distribution and pharmacokinetics of a specific drug to clinical efficacy. Correlation with routine clinical functional measurements is possible, but, whereas 3D imaging data provides local drug deposition information, clinical tests of respiratory status are “black-box” measurements with outcomes specific to large or small airways inferred from the results. However, biopsies may be obtained directly from the tissue being imaged and therefore allow correlations with tracer uptake in the particular tissues. Imaging a radiolabeled pharmaceutic over time provides temporal information of receptor binding, drug absorption, or drug clearance from airways or the alveolar space. Changes in the deposition of inhaled aerosols within the lung related to the presence of disease or resulting from inhalation challenge interventions or inhaled therapies can be visualized with PET and may correlate with clinical outcomes. As well, the amount of an inhaled tracer deposited in various regions of the lung can give an indication of the efficiency of drug delivery and, combined with the regional distribution of the drug within the lung and the rate of drug absorption, estimate clinical efficacy and safety.