ASSESSMENT OF CORONARY HEART DISEASE WITH CARBON DIOXIDE
20220054661 · 2022-02-24
Assignee
Inventors
Cpc classification
A61K33/00
HUMAN NECESSITIES
A61K2300/00
HUMAN NECESSITIES
A61K31/7076
HUMAN NECESSITIES
International classification
Abstract
There are provided methods for diagnosing coronary heart disease in a subject in need thereof comprising administering an admixture comprising CO.sub.2 to a subject to reach a predetermined PaCO.sub.2 in the subject to induce hyperemia, monitoring vascular reactivity in the subject and diagnosing the presence or absence of coronary heart disease in the subject, wherein decreased vascular reactivity in the subject compared to a control subject is indicative of coronary heart disease. There are also provided methods for increasing sensitivity and specificity of BOLD MRI.
Claims
1. A method of inducing hyperemia to diagnose coronary heart disease in a subject in need thereof, comprising: administering a CO.sub.2-containing gas to the subject; attaining at least one increase in the subject's coronary PaCO.sub.2 sufficient for diagnosing coronary heart disease from imaging data; and imaging the heart for a number of repeated measurements during a period in which the at least one increase in PaCO.sub.2 is measurable, to produce imaging data indicative of a cardiovascular-disease-associated vasoreactive response in at least one coronary blood vessel or region of the heart, wherein the number of repeated measurements is sufficient to produce a statistical significance of p<0.05 in measuring a change from baseline in vasoreactive response to the at least one increase in coronary PaCO.sub.2.
2. The method of claim 1, comprising attaining the at least one increase in PaCO.sub.2 in a stepwise manner.
3. The method of claim 1 comprising attaining the at least one increase in PaCO.sub.2 in a block manner.
4. The method of claim 1, comprising administering CO.sub.2 via inhalation to attain a predetermined PaCO.sub.2.
5. The method of claim 4, wherein attaining at least one increase in the subject's coronary PaCO.sub.2 comprises alternating between two or more coronary PaCO.sub.2 levels.
6. The method of claim 5, wherein the predetermined increase in PaCO.sub.2 is an about 8 mmHg to about 20 mmHg increase in a subject's baseline, wherein the baseline is a steady state level measured prior to changing the subject's PaCO.sub.2.
7. The method of claim 1, wherein the cardiovascular disease-associated vasoreactive response is a compromised increase in blood flow.
8. The method of claim 1, wherein the imaging method is PET or SPECT and the measure of the cardiovascular-disease-associated vasoreactive response is the presence or absence of a threshold increase in blood flow.
9. The method of claim 1, the imaging data is indicative of the presence or absence of a twofold increase in blood flow.
10. (canceled)
11. The method of claim 1, wherein the imaging data are obtained by MRI.
12. The method of claim 1, wherein the imaging data are a change in signal intensity of a BOLD MRI signal.
13. (canceled)
14. (canceled)
15. The method of claim 12, wherein the number of repeated measurements includes at least 8 measurements in measuring a 10% increase in the BOLD Mill signal.
16. The method of claim 1, wherein at least one increase in PaCO.sub.2 is at least a 10 mm Hg increase from a first level which is determined to be between 30 and 55 mm Hg.
17. The method of claim 1, wherein at least one increase in PaCO.sub.2 is at least a 10 mm Hg increase from a first level determined to be between 35 and 45 mm Hg.
18. (canceled)
19. (canceled)
20. The method of claim 2, wherein administration of the carbon dioxide containing gas is controlled so as to change the PaCO.sub.2 in 2 to 4 mmHg increments starting from a first value in the range of about 30 mm Hg to 50 mmHg CO.sub.2 and ending a second value which is at least 8-14 mm higher if the increment is 2 mm Hg and at least 10-20 mm Hg higher if the increment is 3-4 mm Hg higher.
21. (canceled)
22. (canceled)
23. A method for imaging hyperemia in a subject in need of a diagnostic assessment of cardiovascular disease comprising administering a CO.sub.2 containing gas in a non-therapeutic diagnostic setting, attaining at least one selected increase in a subject's coronary PaCO.sub.2 sufficient for diagnosing coronary heart disease from imaging data, and imaging the heart for a number of repeated measurements during a period in which the selected increase in PaCO.sub.2 is measurable, wherein the imaging data is selected to be indicative of a cardiovascular-disease-associated vasoreactive response in at least one coronary blood vessel or region of the heart, wherein the number of repeated measurements is sufficient to produce a statistical significance of p<0.05 in measuring a change from baseline in vasoreactive response to the at least one increase in coronary PaCO.sub.2.
24-29. (canceled)
30. The method of claim 23, wherein cardiovascular-disease-associated vasoreactive response is obtained by controlling the administration of a CO.sub.2 containing gas to repeatedly alternate between at least two PaCO.sub.2 levels and obtaining repeat BOLD MRI measurement at each level to statistically assess the vasoreactive response.
31-39. (canceled)
40. A method for controlling a gas flow controller during a cardiac imaging procedure, the gas flow controller operable to deliver controlled amounts of carbon dioxide for inspiration by a subject during free breathing, the method comprising operating the gas mass flow controller to administer controlled amounts of carbon dioxide to attain an at least one altered level of carbon dioxide in the subject's arterial blood, wherein the at least one altered level of carbon dioxide is selected to induce a selected hyperemic response in the subject's myocardium over a time period selected for repeated imaging of the selected hyperemic response, wherein the selected hyperemic response identifies at least one segment of the subject's myocardium with a reduced hyperemic response relative to the selected hyperemic response, and the repeated imaging produces a statistical significance of p<0.05 in the difference between the reduced hyperemic response and the selected hyperemic response.
41. The method of claim 40, wherein the reduced hyperemic response is a decrease in a ratio of myocardial perfusion at stress to myocardial perfusion at rest to 2:1.
42-53. (canceled)
54. The method of claim 40, wherein the altered level of carbon dioxide is a 25 mm of Hg increase from a previous level, optionally a measured baseline level for the subject, optionally a baseline level for the subject at rest, optionally a baseline level for the subject when the subject is breathing at a regulated elevated minute volume.
55-74. (canceled)
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0073] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.
[0074] For a better understanding of the present invention, as well as other aspects and further features thereof, reference is made to the following description which is to be used in conjunction with the accompanying drawings, where:
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DETAILED DESCRIPTION
[0095] All references cited herein are incorporated by reference in their entirety as though fully set forth. Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Singleton et al., Dictionary of Microbiology and Molecular Biology 3.sup.rd ed., J. Wiley & Sons (New York, N.Y. 2001); March, Advanced Organic Chemistry Reactions, Mechanisms and Structure 5.sup.th ed., J. Wiley & Sons (New York, N.Y. 2001); and Sambrook and Russel, Molecular Cloning: A Laboratory Manual 3rd ed., Cold Spring Harbor Laboratory Press (Cold Spring Harbor, N.Y. 2001), provide one skilled in the art with a general guide to many of the terms used in the present application.
[0096] One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present invention. Indeed, the present invention is in no way limited to the methods and materials described. For purposes of the present invention, the following terms are defined below.
[0097] “Beneficial results” may include, but are in no way limited to, lessening or alleviating the severity of the disease condition, preventing the disease condition from worsening, curing the disease condition, preventing the disease condition from developing, lowering the chances of a patient developing the disease condition and prolonging a patient's life or life expectancy.
[0098] “Mammal” as used herein refers to any member of the class Mammalia, including, without limitation, humans and nonhuman primates such as chimpanzees and other apes and monkey species; farm animals such as cattle, sheep, pigs, goats and horses; domestic mammals such as dogs and cats; laboratory animals including rodents such as mice, rats and guinea pigs, and the like. The term does not denote a particular age or sex. Thus, adult and newborn subjects, as well as fetuses, whether male or female, are intended to be included within the scope of this term.
[0099] “Treatment” and “treating,” as used herein refer to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) the targeted pathologic condition, prevent the pathologic condition, pursue or obtain beneficial results, or lower the chances of the individual developing the condition even if the treatment is ultimately unsuccessful. Those in need of treatment include those already with the condition as well as those prone to have the condition or those in whom the condition is to be prevented.
[0100] “Carbogen” as used herein is an admixture of carbon dioxide and oxygen. The amounts of carbon dioxide and oxygen in the admixture may be determined by one skilled in the art.
[0101] Medical grade carbogen is typically 5% CO.sub.2 and 95% O.sub.2. In various other embodiments, carbon dioxide used to induce hyperemia may be an admixture of ranges including but not limited to 94% O.sub.2 and 6% CO.sub.2, 93% O.sub.2 and 7% CO.sub.2, 92% O.sub.2 and 8% CO.sub.2, 91% O.sub.2 and 9% CO.sub.2, 90% O.sub.2 and 10% CO.sub.2, 85% O.sub.2 and 15% CO.sub.2, 80% O.sub.2 and 20% CO.sub.2, 75% O.sub.2 and 25% CO.sub.2 and/or 70% O.sub.2 and 30% CO.sub.2. Optionally, for blending with air, the CO.sub.2 containing gas comprises 20% oxygen.
[0102] “BOLD” as used herein refers to blood-oxygen-level dependence.
[0103] The term “about” is used herein to indicate that a value includes an inherent variation of error for the device or the method being employed to determine the value.
[0104] A “vascular-disease-associated” coronary vasoreactive response means a type and/or quantum of vasoreactive response elicited by cardiac stress testing (e.g. exercise or administration of a hyperemic drug or a CO.sub.2 containing gas) as demonstrable in an imaging study using one or more diagnostic imaging parameters of the type suitable to diagnose coronary vascular disease. For example, with respect to PET and SPECT, a normal response would be considered a four to five-fold increase in blood flow. With respect to BOLD MRI imaging, a 10-12+% increase in BOLD signal would be considered normal. Disease associated responses are those which are not normal in varying significant degrees among which, as evidence of disease, benchmarks may be adopted to categorize differences which represent a clearer-cut diagnosis or a progression of disease that warrants greater follow-up or more proactive treatment, for example a less than two-fold increase in blood flow as measured by PET or SPECT (typically measured in ml. of blood/min/gm of tissue). Accordingly, a benchmark which represents a change from a value that clinicians described as “normal” which is at least statistically significant and optionally is also comparable to a standard for cardiac stress testing adopted by clinicians with respect to inducing stress represents a clear-cut benchmark for using CO.sub.2 as a vasoactive stress stimulus.
[0105] A targeted increase in PaCO.sub.2 will be selected to cause a similar vasoreactive response in normal and diseased tissue. From the standpoint of statistical significance, it will be appreciated that selection of a discriminatory increase in PaCO.sub.2 may depend on whether or not repeat measurements are made, for example, the number of repeat measurements of a BOLD signal intensity that are made while at lower and increased PaCO.sub.2 levels.
[0106] Current methods for inducing hyperemia in subjects include the use of compounds such as adenosine, analogs thereof and/or functional equivalents thereof. However, such compounds (for example, adenosine) have adverse side effects including bradycardia, arrhythmia, transient or prolonged episode of asystole, ventricular fibrillation (rarely), chest pain, headache, dyspnea, and nausea, making it less than favorable for initial or follow-up studies.
[0107] The technology described herein is directed to the use of CO.sub.2 instead of hyperemia-inducing drugs, in view of their side effects, to assess myocardial response and risk of coronary artery diseases. To date, however, it has not been possible to independently control arterial CO.sub.2 and O.sub.2, hence direct association of the influence of partial pressure of CO.sub.2 (PaCO.sub.2) on coronary vasodilation has been difficult to determine. With the development of gas flow controller devices designed to control gas concentrations in the lungs and blood (for example, RespirACT™, Thornhill Research, WO/2013/0082703), it is now possible to precisely control the arterial CO.sub.2, while, in some embodiments, holding O.sub.2 constant. With such devices, the desired PaCO.sub.2 changes are rapid (1-2 breaths) and are independent of minute ventilation. The inventors are among the first adopters of such devices for the assessment of myocardial response to CO.sub.2.
[0108] The methods provided herein are believed to be the first to use modulation of CO.sub.2 levels to show that the carbon dioxide can have the same effect as the clinical dose of other hyperemia-inducing drugs such as adenosine but without the side effects. We report herein that hyperemia is induced by administering an admixture comprising a predetermined amount of CO.sub.2 to a subject in need thereof to assess myocardial response, evaluate coronary artery disease and identify ischemic heart disease. In an embodiment, hyperemia is induced by independently altering the administered CO.sub.2 level while holding oxygen (O.sub.2) constant to assess myocardial response, evaluate coronary artery disease and identify ischemic heart disease. A subject's myocardial response after administration of CO.sub.2 may be monitored using various imaging techniques such as MRI.
[0109] Cardiac Stress Testing
[0110] When exercise stress testing is contra-indicated (in nearly 50% of patients), existing imaging modalities use adenosine (or its analogues such as dipyridamole or regadenoson) to induce hyperemia. However, as described above, adenosine or analogs thereof or functional equivalents thereof, are well known for their adverse side effects such as bradycardia, arrhythmia, transient or prolonged episode of asystole, ventricular fibrillation (rarely), chest pain, headache, dyspnea, and nausea, making it less than favorable for initial or follow-up studies. Direct measures of ischemic burden may be determined on the basis of single-photon emission computed tomography (SPECT/SPET), positron emission tomography (PET), myocardial contrast echocardiography (MCE), and first-pass perfusion magnetic resonance imaging (FPP-Mill). SPECT and PET use radiotracers as contrast agents. While SPECT and PET studies account for approximately 90% of myocardial ischemia-testing studies, the sensitivity and specificity for both methods combined for the determination of severe ischemia is below 70%. Both MCE and FPP-MRI are relatively newer approaches that require the use of exogenous contrast media and intravenous pharmacological stress agent (such as adenosine), both carrying significant risks and side effects in certain patient populations.
[0111] BOLD-MRI
[0112] An alternate method, BOLD (Blood-Oxygen-Level-Dependent) MRI, relies on endogenous contrast mechanisms (changes in blood oxygen saturation, % O.sub.2) to identify ischemic territories. The potential benefits of BOLD MRI for detecting global or regional myocardial ischemia due to coronary artery disease (CAD) were demonstrated at least a decade ago. Although a number of pilot clinical studies have demonstrated the feasibility of using BOLD Mill for identifying clinically significant myocardial ischemia due to CAD, the method is inherently limited by sensitivity and specificity due to low BOLD contrast-to-noise ratio (CNR). In some embodiments of methods provided herein, the repeatability of BOLD MRI using CO.sub.2 provides the means to improve sensitivity and specificity, which is not possible using adenosine or analogs thereof.
[0113] In some embodiments, there is provided a method for increasing the sensitivity and specificity of BOLD MRI. The method includes administering an admixture comprising CO.sub.2 to a subject in need thereof to induce hyperemia and imaging the myocardium using Mill to assess a hypermic response in response to a predetermined modulation in PaCO.sub.2. In some embodiments, this method utilizes (i) an individualized targeted change in arterial partial pressure of CO.sub.2 (PaCO.sub.2) as the non-invasive vasoactive stimulus, (ii) fast, high-resolution, 4D BOLD Mill at 3T and (iii) statistical models (for example, the generalized linear model (GLM) theory) to derive statistical parametric maps (SPM) to reliably detect and quantify the prognostically significant ischemic burden through repeated measurements (i.e., in a data-driven fashion). In some embodiments, the method for increasing the sensitivity and specificity of BOLD MRI comprises (i) obtaining free-breathing cardiac phase-resolved 3D myocardial BOLD images (under different PaCO.sub.2 states established via inhalation of an admixture of gases comprising of CO.sub.2); (ii) registering and segmenting the images to obtain the myocardial dynamic volume; and (iii) identifying ischemic territory and quantifying image volume.
[0114] Obtaining the Images
[0115] The first step in increasing the sensitivity and specificity of BOLD MRI is to obtain free-breathing cardiac phase resolved 3D myocardial BOLD images. Subjects are placed on the MRI scanner table, ECG leads are placed, and necessary surface coils are positioned. Subsequently the subjects' hearts are localized and the cardiac shim protocol is prescribed over the whole heart. K-space lines, time stamped for trigger time are collected using cine SSFP acquisition with image acceleration along the long axis. Central k-space lines corresponding to each cardiac phase will be used to derive the center of mass (COM) curves along the z-axis via 1-D fast Fourier transform (FFT). Based on the COM curves, the k-space lines from each cardiac phase will be sorted into 1-30 bins, each corresponding to a respiratory state with the first bin being the reference bin (end-expiration) and the last bin corresponding to end inspiration.
[0116] To minimize the artifacts from under sampling, the data will be processed with a 3D filter, followed by re-gridding the k-space lines, application of a spatial mask (to restrict the registration to region of the heart) and performing FFT to obtain the under sampled 3D image for each respiratory bin. Using the end-expiration image as the reference image, images from all bins (except bin 1) are registered using kits such as Insight Tool Kit (freely available from www.itk.org), or an equivalent software platform, in an iterative fashion and the transform parameters will be estimated for rotation, scaling, shearing, and translation of heart between the different respiratory bins. The k-space data will again be divided into 1 to 30 respiratory bins, re-gridded, transformed to the reference image (3D affine transform), summed together, and the final 3D image will be reconstructed. Imaging parameters may be TR=3.0 to 10 ms and flip angle=1° to 90°. In this fashion, 3D cine data under controlled PaCO.sub.2 values (hypo- and hyper-carbic states) are collected.
[0117] Registration and Segmentation of Images
[0118] The next step in increasing the sensitivity and specificity of BOLD MRI is registration and segmentation of the images to obtain the myocardial dynamic volume. The pipeline utilizes MATLAB and C++ using the ITK framework or an equivalent software platform. The myocardial MR images obtained with repeat CO.sub.2 stimulation blocks will be loaded in MATLAB (or an equivalent image processing platform) and arranged in a four-dimensional (4D) matrix, where the first 3 dimensions represent volume (voxels) and the fourth dimension is time (cardiac phase). Subsequently, each volume is resampled to achieve isotropic voxel size. End-systole (ES) are identified for each stack based on our minimum cross-correlation approach. A 4D non-linear registration algorithm is used to find voxel-to-voxel correspondences (deformation fields) across all cardiac phases. Using the recovered deformation, all cardiac phases are wrapped to the space of ES, such that all phases are aligned to ES. The next step is to recover the transformations across all ES images from repeat CO.sub.2 blocks and bring them to the same space using a diffeomorphic volume registration tool, such as ANTs. Upon completion, all cardiac phases from all acquisitions will be spatially aligned to the space of ES of the first acquisition (used as reference) and all phase-to-phase deformations and acquisition-to-acquisition transformations will be known. An expert delineation of the myocardium in the ES of the first (reference) acquisition will then be performed. Based on the estimated deformation fields and transformations, this segmentation is propagated to all phases and acquisitions, resulting in fully registered and segmented myocardial dynamic volumes.
[0119] Image Analysis to Identify and Quantify Ischemic Territories
[0120] The final step needed for increasing the sensitivity and specificity of BOLD MRI is identifying ischemic territory and quantifying image volume. Since BOLD responses are optimally observed in systolic frames, only L systolic cardiac volumes (centered at ES) are retained from each fully registered and segmented 4D BOLD MR image set obtained above. Only those voxels contained in the myocardium are retained and the corresponding RPP (rate-pressure-product) and PaCO.sub.2 are noted. Assuming N acquisitions per CO.sub.2 state (hypocarbic or hypercarbic) and K, CO.sub.2 stimulation blocks, and each cardiac volume consists of n×m×p (x=multiplication) isotropic voxels, build a concatenated fully registered 4D dataset consisting of n×m×p×t pixels, where x=multiplication and t=L×K×N, and export this dataset in NIFTI (or an equivalent) format using standard tools. The 4D dataset is loaded into a voxel-based statistical model fitting (such as FSL-FEAT developed for fMRI), to fit the model for each voxel. The statistical analysis outputs a P-statistic volume, i.e., the SPM, where for each voxel in the myocardium the p-value of the significance of the correlation to the model is reported. The statistical parametric maps (SPM) are thresholded by identifying the voxels that have p<0.05. Those voxels are identified as hyperemic for responding to the CO.sub.2 stimulation. The total number of hyperemic voxels (V.sub.H) are counted and their relative volume (V.sub.RH=V.sub.H/total voxels in myocardium) is determined. The voxels that do not respond to CO.sub.2 stimulation (on SPM) are identified as ischemic and used to generate a binary 3D map of ischemic voxels (3D-ISCH.sub.map). In addition, total ischemic voxels (V.sub.I) and the relative ischemic volume (V.sub.RI=V.sub.t/total myocardial voxels) are determined.
[0121] The above methods provide ischemic volumes that can be reliably identified on the basis of statistical analysis applied to repeatedly acquire 4D BOLD images under precisely targeted changes in PaCO.sub.2. These volumes are closely related to the clinical index of fractional flow reserve FFR.
[0122] FFR
[0123] An additional method, fractional flow reserve (FFR) is used in coronary catheterization to measure pressure differences across a coronary artery stenosis to determine the likelihood that the stenosis impedes oxygen delivery to the heart muscle (myocardial ischemia). Fractional flow reserve measures the pressure behind (distal to) a steno sis relative to the pressure before the stenosis, using adenosine or papaverine to induce hyperemia. A cut-off point of 0.75 to 0.80 has been used wherein higher values indicate a non-significant stenosis and lower values indicate a significant lesion. FFR, determined as the relative pressure differences across the stenotic coronary artery has emerged as the new standard for determining clinically significant ischemia (FFR≤0.75). However, it is invasive, expensive, and exposes the patient to ionizing radiation and the side-effects of the use of adenosine. In view of the side-effects of adenosine discussed above, Applicants propose using carbon dioxide instead of adenosine to induce hyperemia, by administering to a subject an admixture comprising CO.sub.2 to reach a predetermined PaCO.sub.2 in the subject to induce hyperemia. In some embodiments, the admixture comprises any one or more of carbon dioxide, oxygen, and nitrogen, e.g., carbon dioxide, oxygen and nitrogen; carbon dioxide and oxygen; carbon dioxide and nitrogen; or carbon dioxide alone. In one embodiment, the amounts of CO.sub.2 and O.sub.2 administered are both altered. In another embodiment, the amount of CO.sub.2 administered is altered to a predetermined level while the amount of O.sub.2 administered is held constant. In various embodiments, the amounts of any one or more of CO.sub.2, O.sub.2 and N2 in an admixture are changed or held constant as would be readily apparent to a person having ordinary skill in the art.
[0124] Methods
[0125] There are provided herein methods for diagnosing coronary heart disease in a subject in need thereof comprising administering an admixture comprising CO.sub.2 to a subject to reach a predetermined PaCO.sub.2 in the subject to induce hyperemia, monitoring vascular reactivity in the subject and diagnosing the presence or absence of coronary heart disease in the subject, wherein decreased vascular reactivity in the subject compared to a control subject is indicative of coronary heart disease. In an embodiment, CO.sub.2 is administered via inhalation. In another embodiment, CO.sub.2 levels are altered while the O.sub.2 levels remain unchanged so that the PaCO.sub.2 is changed independently of the O.sub.2 level. In a further embodiment, vascular reactivity is monitored using imagining techniques deemed appropriate by one skilled in the art, including but not limited to any one or more of positron emission tomography (PET), single photon emission computed tomography/computed tomography (SPECT), computed tomography (CT), magnetic resonance imaging (MRI), functional magnetic resonance imaging (fMRT), single photon emission computed tomography/computed tomography (SPECT/CT), positron emission tomography/computed tomography (PET/CT), ultrasound, electrocardiogram (ECG), Electron-beam computed tomography (EBCT), echocardiogram (ECHO), and electron spin resonance (ESR), and/or any combination of the imaging modalities such as (PET/MR), PET/CT, and/or SPECT/MR. In an embodiment, vascular reactivity is monitored using free-breathing BOLD MRI. In some embodiments, the admixture comprises any one or more of carbon dioxide, oxygen and nitrogen, e.g., carbon dioxide, oxygen and nitrogen; carbon dioxide and oxygen; carbon dioxide and nitrogen; or carbon dioxide alone. In one embodiment, the amounts of CO.sub.2 and O.sub.2 administered are both altered. In another embodiment, the amount of CO.sub.2 administered is altered to a predetermined level while the amount of O.sub.2 administered is held constant. In various embodiments, the amounts of any one or more of CO.sub.2, O.sub.2 and N.sub.2 in an admixture are changed or held constant as would be readily apparent to a person having ordinary skill in the art.
[0126] The invention also provides a method for assessing hyperemic response in a subject in need thereof comprising administering an admixture comprising CO.sub.2 to a subject to reach a predetermined PaCO.sub.2 in the subject to induce hyperemia, monitoring vascular reactivity in the subject and assessing hyperemic response in the subject, wherein decreased vascular reactivity in the subject compared to a control subject is indicative of poor hyperemic response, thereby assessing hyperemic response in the subject in need thereof. This method may also be used to assess organ perfusion and assess vascular reactivity. In an embodiment, CO.sub.2 is administered via inhalation. In another embodiment, CO.sub.2 levels are altered while the O.sub.2 levels remain unchanged so that the PaCO.sub.2 is changed independently of the O.sub.2 level. In a further embodiment, vascular reactivity is monitored using imagining techniques deemed appropriate by one skilled in the art, including but not limited to any one or more of positron emission tomography (PET), single photon emission computed tomography/computed tomography (SPECT), computed tomography (CT), magnetic resonance imaging (MRI), functional magnetic resonance imaging (fMRI), single photon emission computed tomography/computed tomography (SPECT/CT), positron emission tomography/computed tomography (PET/CT), ultrasound, electrocardiogram (ECG), Electron-beam computed tomography (EBCT), echocardiogram (ECHO), and electron spin resonance (ESR), and/or any combination of the imaging modalities such as (PET/MR), PET/CT, and/or SPECT/MR. In an embodiment, vascular reactivity is monitored using free-breathing BOLD MRI. In some embodiments, the admixture comprises any one or more of carbon dioxide, oxygen and nitrogen, e.g., carbon dioxide, oxygen and nitrogen; carbon dioxide and oxygen; carbon dioxide and nitrogen; or carbon dioxide alone. In one embodiment, the amounts of CO.sub.2 and O.sub.2 administered are both altered. In another embodiment, the amount of CO.sub.2 administered is altered to a predetermined level while the amount of O.sub.2 administered is held constant. In various embodiments, the amounts of any one or more of CO.sub.2, O.sub.2 and N2 in an admixture are changed or held constant as would be readily apparent to a person having ordinary skill in the art.
[0127] The invention is further directed to methods for producing coronary vasodilation in a subject in need thereof comprising providing a composition comprising CO.sub.2 and administering the composition comprising CO.sub.2 to a subject to reach a predetermined PaCO.sub.2 in the subject so as to produce coronary vasodilation in the subject, thereby producing coronary vasodilation in the subject. In an embodiment, CO.sub.2 is administered via inhalation. In another embodiment, CO.sub.2 levels are altered while the O.sub.2 levels remain unchanged so that the PaCO.sub.2 is changed independently of the O.sub.2 level. In a further embodiment, vascular reactivity is monitored using imagining techniques deemed appropriate by one skilled in the art, including but not limited to any one or more of positron emission tomography (PET), single photon emission computed tomography/computed tomography (SPECT), computed tomography (CT), magnetic resonance imaging (MRI), functional magnetic resonance imaging (fMRI), single photon emission computed tomography/computed tomography (SPECT/CT), positron emission tomography/computed tomography (PET/CT), ultrasound, electrocardiogram (ECG), Electron-beam computed tomography (EBCT), echocardiogram (ECHO), and electron spin resonance (ESR) and/or any combination of the imaging modalities such as (PET/MR), PET/CT, and/or SPECT/MR. In an embodiment, vascular reactivity is monitored using free-breathing BOLD MRI. In some embodiments, the admixture comprises any one or more of carbon dioxide, oxygen and nitrogen, e.g., carbon dioxide, oxygen and nitrogen; carbon dioxide and oxygen; carbon dioxide and nitrogen; or carbon dioxide alone. In one embodiment, the amounts of CO.sub.2 and O.sub.2 administered are both altered. In another embodiment, the amount of CO.sub.2 administered is altered to a predetermined level while the amount of O.sub.2 administered is held constant. In various embodiments, the amounts of any one or more of CO.sub.2, O.sub.2 and N.sub.2 in an admixture are changed or held constant as would be readily apparent to a person having ordinary skill in the art.
[0128] The invention also provides a method for assessing tissue and/or organ perfusion in a subject in need thereof comprising administering an admixture comprising CO.sub.2 to a subject to reach a predetermined PaCO.sub.2 in the subject to induce hyperemia, monitoring vascular reactivity in the tissue and/or organ and assessing tissue and/or organ perfusion by assessing the hyperemic response in the subject, wherein decreased vascular reactivity in the subject compared to a control subject is indicative of poor hyperemic response and therefore poor tissue and/or organ perfusion. In an embodiment, CO.sub.2 is administered via inhalation. In another embodiment, CO.sub.2 levels are altered while the O.sub.2 levels remain unchanged so that the PaCO.sub.2 is changed independently of the O.sub.2 level. In a further embodiment, vascular reactivity is monitored using imagining techniques deemed appropriate by one skilled in the art, including but not limited to any one or more of positron emission tomography (PET), single photon emission computed tomography/computed tomography (SPECT), computed tomography (CT), magnetic resonance imaging (MRI), functional magnetic resonance imaging (fMRI), single photon emission computed tomography/computed tomography (SPECT/CT), positron emission tomography/computed tomography (PET/CT), ultrasound, electrocardiogram (ECG), Electron-beam computed tomography (EBCT), echocardiogram (ECHO), and electron spin resonance (ESR), and/or any combination of the imaging modalities such as (PET/MR), PET/CT, and/or SPECT/MR. In an embodiment, vascular reactivity is monitored using free-breathing BOLD MRI. In some embodiments, the admixture comprises any one or more of carbon dioxide, oxygen and nitrogen, e.g., carbon dioxide, oxygen and nitrogen; carbon dioxide and oxygen; carbon dioxide and nitrogen; or carbon dioxide alone. In one embodiment, the amounts of CO.sub.2 and O.sub.2 administered are both altered. In another embodiment, the amount of CO.sub.2 administered is altered to a predetermined level while the amount of O.sub.2 administered is held constant. In various embodiments, the amounts of any one or more of CO.sub.2, O.sub.2 and N.sub.2 in an admixture are changed or held constant as would be readily apparent to a person having ordinary skill in the art.
[0129] In some embodiments, the admixture comprising CO.sub.2 is administered at high doses for short duration or at low doses for longer durations. For example, administering the admixture comprising CO.sub.2 at high doses of CO.sub.2 for a short duration comprises administering any one or more of 40 mmHg to 45 mmHg, 45 mmHg to 50 mmHg, 50 mmHg to 55 mmHg, 55 mmHg CO.sub.2 to 60 mm Hg CO.sub.2, 60 mmHg CO.sub.2 to 65 mm Hg CO.sub.2, 65 mmHg CO.sub.2 to 70 mm Hg CO.sub.2, 70 mmHg CO.sub.2 to 75 mm Hg CO.sub.2, 75 mmHg CO.sub.2 to 80 mm Hg CO.sub.2, 80 mmHg CO.sub.2 to 85 mm Hg CO.sub.2 or a combination thereof, for about 20 minutes, 15 minutes, 10 minutes, 9 minutes, 8 minutes, 7 minutes, 6 minutes, 5 minutes, 4 minutes, 3 minutes, 2 minutes, 1 minute or a combination thereof. In various embodiments, the predetermined levels of CO.sub.2 are administered so that the arterial level of CO.sub.2 reaches the PaCO.sub.2 of any one or more of the above ranges. In an embodiment, the predetermined levels of CO.sub.2 are administered so that the arterial level of CO.sub.2 reaches the PaCO.sub.2 of about 60 mm Hg.
[0130] For example, administering low doses of predetermined amounts of CO.sub.2 for a longer duration comprises administering the predetermined amount of CO.sub.2 at any one or more of about 30 mmHg CO.sub.2 to about 35 mmHg CO.sub.2, about 35 mmHg CO.sub.2 to about 40 mmHg CO.sub.2, about 40 mmHg CO.sub.2 to about 45 mmHg CO.sub.2, about 60 mmHg CO.sub.2 to about 65 mmHg CO.sub.2, or a combination thereof for any one or more of about 20 to 24 hours, about 15 to 20 hours, about 10 to 15 hours, about 5 to 10 hours, about 4 to 5 hours, about 3 to 4 hours, about 2 to 3 hours, and about 1 to 2 hours, or a combination thereof, before inducing hyperemia. In various embodiments, the predetermined levels of CO.sub.2 are administered so that the arterial level of CO.sub.2 reaches the PaCO.sub.2 of any one or more of the above ranges.
[0131] In still further embodiments, the predetermined levels of CO.sub.2 are administered so that the arterial level of CO.sub.2 reaches a PaCO.sub.2 that is increased by about 25 mmHg in a subject, i.e., an about 25 mm Hg increase in PaCO.sub.2 is achieved in the subject after the CO.sub.2 administration. In an embodiment, the predetermined levels of CO.sub.2 are administered so that the arterial level of CO.sub.2 reaches the PaCO.sub.2 of about 60 mm Hg. For example, PaCO.sub.2 may be altered from a baseline level of about 35 mm Hg to about 60 mm Hg in the subject. In some embodiments, the PaCO.sub.2 is increased by about 22 mm Hg to about 28 mm Hg in the subject, or by about 22 mm Hg, about 23 mm Hg, about 24 mm Hg, about 25 mm Hg, about 26 mm Hg, about 27 mm Hg, or about 28 mm Hg in the subject.
[0132] In some embodiments, the predetermined levels of CO.sub.2 are administered so that the hyperemic response in the subject is an about two-fold increase in the subject's myocardial blood flow relative to a measured baseline value in the subject (e.g., value before administration of CO.sub.2).
[0133] In some embodiments, the predetermined levels of CO.sub.2 are administered so that the hyperemic response in the subject is substantially the same as the hyperemic response obtained using a hyperemia-inducing drug such as adenosine, e.g., is substantially the same as a reference standard. In some embodiments, the predetermined levels of CO.sub.2 are selected to induce a selected hyperemic response in the subject. In some embodiments, the selected hyperemic response is a reference standard hyperemic response. In some embodiments, the selected hyperemic response is a hyperemic response that is sufficient to show a response deficit in ischemic tissue, i.e. sufficient to enable imaging of a reduced hyperemic response in ischemic tissue in at least one segment of the subject's myocardium in a cardiac imaging procedure.
[0134] In one embodiment, CO.sub.2 is administered in a stepwise manner. In another embodiment, administering carbon dioxide in a stepwise manner includes administering carbon dioxide in 5 mmHg increments in the range of any one or more of 10 mmHg to 100 mmHg CO.sub.2, 20 mmHg to 100 mmHg CO.sub.2, 30 mmHg to 100 mmHg CO.sub.2, 40 mmHg to 100 mmHg CO.sub.2, 50 mmHg to 100 mmHg CO.sub.2, 60 mmHg to 100 mmHg CO.sub.2, 10 mmHg to 90 mmHg CO.sub.2, 20 mmHg to 90 mmHg CO.sub.2, 30 mmHg to 90 mmHg CO.sub.2, 40 mmHg to 90 mmHg CO.sub.2, 50 mmHg to 90 mmHg CO.sub.2, 60 mmHg to 90 mmHg CO.sub.2, 10 mmHg to 80 mmHg CO.sub.2, 20 mmHg to 80 mmHg CO.sub.2, 30 mmHg to 80 mmHg CO.sub.2, 40 mmHg to 80 mmHg CO.sub.2, 50 mmHg to 80 mmHg CO.sub.2, 60 mmHg to 80 mmHg CO.sub.2, 10 mmHg to 70 mmHg CO.sub.2, 20 mmHg to 70 mmHg CO.sub.2, 30 mmHg to 70 mmHg CO.sub.2, 40 mmHg to 70 mmHg CO.sub.2, 50 mmHg to 70 mmHg CO.sub.2, 60 mmHg to 70 mmHg CO.sub.2, 10 mmHg to 60 mmHg CO.sub.2, 20 mmHg to 70 mmHg CO.sub.2, 30 mmHg to 70 mmHg CO.sub.2, 40 mmHg to 70 mmHg CO.sub.2, 50 mmHg to 70 mmHg CO.sub.2, 60 mmHg to 70 mmHg CO.sub.2, 10 mmHg to 60 mmHg CO.sub.2, 20 mmHg to 60 mmHg CO.sub.2, 30 mmHg to 60 mmHg CO.sub.2, 40 mmHg to 60 mmHg CO.sub.2 and 50 mmHg to 60 mmHg CO.sub.2. In various embodiments, the predetermined levels of CO.sub.2 are administered so that the arterial level of CO.sub.2 reaches the PaCO.sub.2 of any one or more of the above ranges.
[0135] In another embodiment, administering carbon dioxide in a stepwise manner includes administering carbon dioxide in 10 mmHg increments in the range of any one or more of 10 mmHg to 100 mmHg CO.sub.2, 20 mmHg to 100 mmHg CO.sub.2, 30 mmHg to 100 mmHg CO.sub.2, 40 mmHg to 100 mmHg CO.sub.2, 50 mmHg to 100 mmHg CO.sub.2, 60 mmHg to 100 mmHg CO.sub.2, 10 mmHg to 90 mmHg CO.sub.2, 20 mmHg to 90 mmHg CO.sub.2, 30 mmHg to 90 mmHg CO.sub.2, 40 mmHg to 90 mmHg CO.sub.2, 50 mmHg to 90 mmHg CO.sub.2, 60 mmHg to 90 mmHg CO.sub.2, 10 mmHg to 80 mmHg CO.sub.2, 20 mmHg to 80 mmHg CO.sub.2, 30 mmHg to 80 mmHg CO.sub.2, 40 mmHg to 80 mmHg CO.sub.2, 50 mmHg to 80 mmHg CO.sub.2, 60 mmHg to 80 mmHg CO.sub.2, 10 mmHg to 70 mmHg CO.sub.2, 20 mmHg to 70 mmHg CO.sub.2, 30 mmHg to 70 mmHg CO.sub.2, 40 mmHg to 70 mmHg CO.sub.2, 50 mmHg to 70 mmHg CO.sub.2, 60 mmHg to 70 mmHg CO.sub.2, 10 mmHg to 60 mmHg CO.sub.2, 20 mmHg to 70 mmHg CO.sub.2, 30 mmHg to 70 mmHg CO.sub.2, 40 mmHg to 70 mmHg CO.sub.2, 50 mmHg to 70 mmHg CO.sub.2, 60 mmHg to 70 mmHg CO.sub.2, 10 mmHg to 60 mmHg CO.sub.2, 20 mmHg to 60 mmHg CO.sub.2, 30 mmHg to 60 mmHg CO.sub.2, 40 mmHg to 60 mmHg CO.sub.2 and 50 mmHg to 60 mmHg CO.sub.2. In various embodiments, the predetermined levels of CO.sub.2 are administered so that the arterial level of CO.sub.2 reaches the PaCO.sub.2 of any one or more of the above ranges.
[0136] In a further embodiment, administering carbon dioxide in a stepwise manner includes administering carbon dioxide in 20 mmHg increments in the range of any one or more of 10 mmHg to 100 mmHg CO.sub.2, 20 mmHg to 100 mmHg CO.sub.2, 30 mmHg to 100 mmHg CO.sub.2, 40 mmHg to 100 mmHg CO.sub.2, 50 mmHg to 100 mmHg CO.sub.2, 60 mmHg to 100 mmHg CO.sub.2, 10 mmHg to 90 mmHg CO.sub.2, 20 mmHg to 90 mmHg CO.sub.2, 30 mmHg to 90 mmHg CO.sub.2, 40 mmHg to 90 mmHg CO.sub.2, 50 mmHg to 90 mmHg CO.sub.2, 60 mmHg to 90 mmHg CO.sub.2, 10 mmHg to 80 mmHg CO.sub.2, 20 mmHg to 80 mmHg CO.sub.2, 30 mmHg to 80 mmHg CO.sub.2, 40 mmHg to 80 mmHg CO.sub.2, 50 mmHg to 80 mmHg CO.sub.2, 60 mmHg to 80 mmHg CO.sub.2, 10 mmHg to 70 mmHg CO.sub.2, 20 mmHg to 70 mmHg CO.sub.2, 30 mmHg to 70 mmHg CO.sub.2, 40 mmHg to 70 mmHg CO.sub.2, 50 mmHg to 70 mmHg CO.sub.2, 60 mmHg to 70 mmHg CO.sub.2, 10 mmHg to 60 mmHg CO.sub.2, 20 mmHg to 70 mmHg CO.sub.2, 30 mmHg to 70 mmHg CO.sub.2, 40 mmHg to 70 mmHg CO.sub.2, 50 mmHg to 70 mmHg CO.sub.2, 60 mmHg to 70 mmHg CO.sub.2, 10 mmHg to 60 mmHg CO.sub.2, 20 mmHg to 60 mmHg CO.sub.2, 30 mmHg to 60 mmHg CO.sub.2, 40 mmHg to 60 mmHg CO.sub.2 and 50 mmHg to 60 mmHg CO.sub.2. In various embodiments, the predetermined levels of CO.sub.2 are administered so that the arterial level of CO.sub.2 reaches the PaCO.sub.2 of any one or more of the above ranges.
[0137] In a further embodiment, administering carbon dioxide in a stepwise manner includes administering carbon dioxide in 25 mmHg increments in the range of any one or more of 10 mmHg to 100 mmHg CO.sub.2, 20 mmHg to 100 mmHg CO.sub.2, 30 mmHg to 100 mmHg CO.sub.2, 40 mmHg to 100 mmHg CO.sub.2, 50 mmHg to 100 mmHg CO.sub.2, 60 mmHg to 100 mmHg CO.sub.2, 10 mmHg to 90 mmHg CO.sub.2, 20 mmHg to 90 mmHg CO.sub.2, 30 mmHg to 90 mmHg CO.sub.2, 40 mmHg to 90 mmHg CO.sub.2, 50 mmHg to 90 mmHg CO.sub.2, 60 mmHg to 90 mmHg CO.sub.2, 10 mmHg to 80 mmHg CO.sub.2, 20 mmHg to 80 mmHg CO.sub.2, 30 mmHg to 80 mmHg CO.sub.2, 40 mmHg to 80 mmHg CO.sub.2, 50 mmHg to 80 mmHg CO.sub.2, 60 mmHg to 80 mmHg CO.sub.2, 10 mmHg to 70 mmHg CO.sub.2, 20 mmHg to 70 mmHg CO.sub.2, 30 mmHg to 70 mmHg CO.sub.2, 40 mmHg to 70 mmHg CO.sub.2, 50 mmHg to 70 mmHg CO.sub.2, 60 mmHg to 70 mmHg CO.sub.2, 10 mmHg to 60 mmHg CO.sub.2, 20 mmHg to 70 mmHg CO.sub.2, 30 mmHg to 70 mmHg CO.sub.2, 40 mmHg to 70 mmHg CO.sub.2, 50 mmHg to 70 mmHg CO.sub.2, 60 mmHg to 70 mmHg CO.sub.2, 10 mmHg to 60 mmHg CO.sub.2, 20 mmHg to 60 mmHg CO.sub.2, 30 mmHg to 60 mmHg CO.sub.2, 40 mmHg to 60 mmHg CO.sub.2 and 50 mmHg to 60 mmHg CO.sub.2. In various embodiments, the predetermined levels of CO.sub.2 are administered so that the arterial level of CO.sub.2 reaches the PaCO.sub.2 of any one or more of the above ranges.
[0138] In a further embodiment, administering carbon dioxide in a stepwise manner includes administering carbon dioxide in 30 mmHg increments in the range of any one or more of 10 mmHg to 100 mmHg CO.sub.2, 20 mmHg to 100 mmHg CO.sub.2, 30 mmHg to 100 mmHg CO.sub.2, 40 mmHg to 100 mmHg CO.sub.2, 50 mmHg to 100 mmHg CO.sub.2, 60 mmHg to 100 mmHg CO.sub.2, 10 mmHg to 90 mmHg CO.sub.2, 20 mmHg to 90 mmHg CO.sub.2, 30 mmHg to 90 mmHg CO.sub.2, 40 mmHg to 90 mmHg CO.sub.2, 50 mmHg to 90 mmHg CO.sub.2, 60 mmHg to 90 mmHg CO.sub.2, 10 mmHg to 80 mmHg CO.sub.2, 20 mmHg to 80 mmHg CO.sub.2, 30 mmHg to 80 mmHg CO.sub.2, 40 mmHg to 80 mmHg CO.sub.2, 50 mmHg to 80 mmHg CO.sub.2, 60 mmHg to 80 mmHg CO.sub.2, 10 mmHg to 70 mmHg CO.sub.2, 20 mmHg to 70 mmHg CO.sub.2, 30 mmHg to 70 mmHg CO.sub.2, 40 mmHg to 70 mmHg CO.sub.2, 50 mmHg to 70 mmHg CO.sub.2, 60 mmHg to 70 mmHg CO.sub.2, 10 mmHg to 60 mmHg CO.sub.2, 20 mmHg to 70 mmHg CO.sub.2, 30 mmHg to 70 mmHg CO.sub.2, 40 mmHg to 70 mmHg CO.sub.2, 50 mmHg to 70 mmHg CO.sub.2, 60 mmHg to 70 mmHg CO.sub.2, 10 mmHg to 60 mmHg CO.sub.2, 20 mmHg to 60 mmHg CO.sub.2, 30 mmHg to 60 mmHg CO.sub.2, 40 mmHg to 60 mmHg CO.sub.2 and 50 mmHg to 60 mmHg CO.sub.2. In various embodiments, the predetermined levels of CO.sub.2 are administered so that the arterial level of CO.sub.2 reaches the PaCO.sub.2 of any one or more of the above ranges.
[0139] In a further embodiment, administering carbon dioxide in a stepwise manner includes administering carbon dioxide in 40 mmHg increments in the range of any one or more of 10 mmHg to 100 mmHg CO.sub.2, 20 mmHg to 100 mmHg CO.sub.2, 30 mmHg to 100 mmHg CO.sub.2, 40 mmHg to 100 mmHg CO.sub.2, 50 mmHg to 100 mmHg CO.sub.2, 60 mmHg to 100 mmHg CO.sub.2, 10 mmHg to 90 mmHg CO.sub.2, 20 mmHg to 90 mmHg CO.sub.2, 30 mmHg to 90 mmHg CO.sub.2, 40 mmHg to 90 mmHg CO.sub.2, 50 mmHg to 90 mmHg CO.sub.2, 60 mmHg to 90 mmHg CO.sub.2, 10 mmHg to 80 mmHg CO.sub.2, 20 mmHg to 80 mmHg CO.sub.2, 30 mmHg to 80 mmHg CO.sub.2, 40 mmHg to 80 mmHg CO.sub.2, 50 mmHg to 80 mmHg CO.sub.2, 60 mmHg to 80 mmHg CO.sub.2, 10 mmHg to 70 mmHg CO.sub.2, 20 mmHg to 70 mmHg CO.sub.2, 30 mmHg to 70 mmHg CO.sub.2, 40 mmHg to 70 mmHg CO.sub.2, 50 mmHg to 70 mmHg CO.sub.2, 60 mmHg to 70 mmHg CO.sub.2, 10 mmHg to 60 mmHg CO.sub.2, 20 mmHg to 70 mmHg CO.sub.2, 30 mmHg to 70 mmHg CO.sub.2, 40 mmHg to 70 mmHg CO.sub.2, 50 mmHg to 70 mmHg CO.sub.2, 60 mmHg to 70 mmHg CO.sub.2, 10 mmHg to 60 mmHg CO.sub.2, 20 mmHg to 60 mmHg CO.sub.2, 30 mmHg to 60 mmHg CO.sub.2, 40 mmHg to 60 mmHg CO.sub.2 and 50 mmHg to 60 mmHg CO.sub.2. In various embodiments, the predetermined levels of CO.sub.2 are administered so that the arterial level of CO.sub.2 reaches the PaCO.sub.2 of any one or more of the above ranges.
[0140] In a further embodiment, administering carbon dioxide in a stepwise manner includes administering carbon dioxide in 50 mmHg increments in the range of any one or more of 10 mmHg to 100 mmHg CO.sub.2, 20 mmHg to 100 mmHg CO.sub.2, 30 mmHg to 100 mmHg CO.sub.2, 40 mmHg to 100 mmHg CO.sub.2, 50 mmHg to 100 mmHg CO.sub.2, 60 mmHg to 100 mmHg CO.sub.2, 10 mmHg to 90 mmHg CO.sub.2, 20 mmHg to 90 mmHg CO.sub.2, 30 mmHg to 90 mmHg CO.sub.2, 40 mmHg to 90 mmHg CO.sub.2, 50 mmHg to 90 mmHg CO.sub.2, 60 mmHg to 90 mmHg CO.sub.2, 10 mmHg to 80 mmHg CO.sub.2, 20 mmHg to 80 mmHg CO.sub.2, 30 mmHg to 80 mmHg CO.sub.2, 40 mmHg to 80 mmHg CO.sub.2, 50 mmHg to 80 mmHg CO.sub.2, 60 mmHg to 80 mmHg CO.sub.2, 10 mmHg to 70 mmHg CO.sub.2, 20 mmHg to 70 mmHg CO.sub.2, 30 mmHg to 70 mmHg CO.sub.2, 40 mmHg to 70 mmHg CO.sub.2, 50 mmHg to 70 mmHg CO.sub.2, 60 mmHg to 70 mmHg CO.sub.2, 10 mmHg to 60 mmHg CO.sub.2, 20 mmHg to 70 mmHg CO.sub.2, 30 mmHg to 70 mmHg CO.sub.2, 40 mmHg to 70 mmHg CO.sub.2, 50 mmHg to 70 mmHg CO.sub.2, 60 mmHg to 70 mmHg CO.sub.2, 10 mmHg to 60 mmHg CO.sub.2, 20 mmHg to 60 mmHg CO.sub.2, 30 mmHg to 60 mmHg CO.sub.2, 40 mmHg to 60 mmHg CO.sub.2 and 50 mmHg to 60 mmHg CO.sub.2. In various embodiments, the predetermined levels of CO.sub.2 are administered so that the arterial level of CO.sub.2 reaches the PaCO.sub.2 of any one or more of the above ranges.
[0141] Other increments of carbon dioxide to be administered in a stepwise manner will be readily apparent to a person having ordinary skill in the art.
[0142] In a further embodiment, a predetermined amount of CO.sub.2 is administered in a block manner. Block administration of carbon dioxide comprises administering carbon dioxide in alternating amounts over a period of time. “In alternating amounts” of CO.sub.2 comprises alternating between any of 20 mmHg and 40 mmHg, 30 mmHg and 40 mmHg, 20 mmHg and 50 mmHg, 30 mmHg and 50 mmHg, 40 mmHg and 50 mmHg, 20 mmHg and 60 mmHg, 30 mmHg and 60 mmHg, 40 mmHg and 60 mmHg, and 50 mmHg and 60 mmHg. In various embodiments, the predetermined levels of CO.sub.2 are administered so that the arterial level of CO.sub.2 reaches the PaCO.sub.2 of any one or more of the above ranges. Other amounts of carbon dioxide to be used in alternating amounts over a period of time will be readily apparent to a person having ordinary skill in the art.
[0143] In one embodiment, vascular reactivity may be measured by characterization of Myocardial Perfusion Reserve, which is defined as a ratio of Myocardial Perfusion at Stress to Myocardial Perfusion at Rest. In healthy subjects the ratio may vary from 5:1 to 6:1. The ratio diminishes with disease. A decrease in this ratio to 2:1 from the healthy level is considered to be clinically significant and indicative of poor vascular reactivity.
[0144] In another embodiment, vascular reactivity may be measured via differential absolute perfusion, which may be obtained using imaging methods such as first pass perfusion, SPECT/PET, CT perfusion or echocardiography in units of ml/sec/g of tissue.
[0145] In an embodiment the CO.sub.2 gas is administered via inhalation. CO.sub.2 may be administered using, for example, RespirACT™ technology from Thornhill Research. In various embodiments, CO.sub.2 is administered for 1-2 minutes, 2-4 minutes, 4-6 minutes, 6-8 minutes, 8-10 minutes, 10-12 minutes, 12-14 minutes, 14-16 minutes, 16-18 minutes and/or 18-20 minutes. In one embodiment, CO.sub.2 is administered for 4-6 minutes. In an additional embodiment, CO.sub.2 is administered for an amount of time it takes for the arterial PaCO.sub.2 (partial pressure of carbon dioxide) to reach 50-60 mmHg from the standard levels of 30 mmHg during CO.sub.2-enhanced imaging.
[0146] In one embodiment, carbon dioxide used to induce hyperemia is medical-grade carbogen which is an admixture of 95% O.sub.2 and 5% CO.sub.2. In various other embodiments, carbon dioxide used to induce hyperemia may be an admixture of ranges including but not limited to 94% O.sub.2 and 6% CO.sub.2, 93% O.sub.2 and 7% CO.sub.2, 92% O.sub.2 and 8% CO.sub.2, 91% O.sub.2 and 9% CO.sub.2, 90% O.sub.2 and 10% CO.sub.2, 85% O.sub.2 and 15% CO.sub.2, 80% O.sub.2 and 20% CO.sub.2, 75% O.sub.2 and 25% CO.sub.2 and/or 70% O.sub.2 and 30% CO.sub.2.
[0147] In another embodiment, vascular reactivity and/or vasodilation are monitored using any one or more of positron emission tomography (PET), single photon emission computed tomography/computed tomography (SPECT), computed tomography (CT), magnetic resonance imaging (MRI), functional magnetic resonance imaging (fMRI), single photon emission computed tomography/computed tomography (SPECT/CT), positron emission tomography/computed tomography (PET/CT), ultrasound, electrocardiogram (ECG), Electron-beam computed tomography (EBCT), echocardiogram (ECHO), and electron spin resonance (ESR), and/or any combination of the imaging modalities such as (PET/MR), PET/CT, and/or SPECT/MR In an embodiment, vascular reactivity is monitored using free-breathing BOLD MRI.
[0148] Imaging techniques using carbon dioxide involve a free-breathing approach so as to permit entry of CO.sub.2 into the subject's system. In an embodiment, the subjects include mammalian subjects, including, human, monkey, ape, dog, cat, cow, horse, goat, pig, rabbit, mouse and rat. In a preferred embodiment, the subject is human. It should be noted that the terms “subject” and “patient” are used interchangeably herein.
Advantages of the Invention
[0149] The methods described herein to functionally assess the oxygen status of the myocardium include administering an effective amount of CO.sub.2 to a subject in need thereof. In an embodiment, the O.sub.2 level is held constant while the CO.sub.2 level is altered so as to induce hyperemia. Applicants herein show the vascular reactivity in subjects in response to changes in PaCO.sub.2. The existing methods use adenosine, dipyridamole, or regadenoson which have significant side-effects, as described above. As described in the Examples below, in some embodiments CO.sub.2 is at least as effective as the existing methods (which use, for example, adenosine) but without the side effects. Methods described herein may provide one or more of the following advantages.
[0150] The use of CO.sub.2 can provide distinct advantages over the use of, for example, adenosine. Administering CO.sub.2 is truly non-invasive because it merely involves inhaling physiologically sound levels of CO.sub.2. The instant methods are simple, repeatable and fast and most likely have the best chance for reproducibility. Not even breath-holding is necessary during acquisition of images using the methods described herein. The instant methods can also be highly cost-effective as no pharmacological stress agents are required, cardiologists may not need to be present during imaging and rapid imaging reduces scan times and costs. Further, in some embodiments CO.sub.2 can produce a selected hyperemic response despite consumption of caffeine, which is advantageous compared to some hyperemia-inducing drugs which do not produce reliable assessments in subjects who have consumed caffeine beforehand.
[0151] Further, the improved BOLD MRI technique described above can provide a non-invasive and reliable determination of ischemic volume (no radiation, contrast-media, or adenosine) and other value-added imaging biomarkers from the same acquisition (Ejection Fraction, Wall Thickening). Additionally, the subject does not experience adenosine-related adverse side effects and thus greater patient tolerance for repeat ischemia testing is achieved. In some embodiments, there is a significant cost-savings from abandoning exogenous contrast media and adenosine/regadenoson. Moreover, the proposed BOLD MRI paradigm can be accompanied by significant technical advances as well: (i) a fast, high-resolution, free-breathing 4D SSFP MRI at 3T, that can impact cardiac MRI in general; (ii) Repeated stimulations of the heart via precisely targeted changes in PaCO.sub.2; and (iii) adoption of sophisticated analytical methods employed in the brain to the heart.
EXAMPLES
[0152] In Examples 1-6, all imaging studies were performed in instrumented animals with a Doppler flow probe attached to the LAD coronary arteries for measurement of flow changes in response to CO.sub.2 and clinical dose of adenosine. In these studies, CO.sub.2 and O.sub.2 delivery were tightly controlled using Respiract. CO.sub.2 values were incremented in steps of 10 mmHg starting from 30 mmHg to 60 mmHg and were ramped down in decrements of 10 mmHg. At each CO.sub.2 level, free-breathing and cardiac gated blood-oxygen-level-dependent (BOLD) acquisitions were prescribed at mid diastole and Doppler flow velocities were measured. Similar acquisitions were also performed with block sequences of CO.sub.2 levels; that is, CO.sub.2 levels were alternated between 40 and 50 mmHg and BOLD images (and corresponding Doppler flow velocities) were acquired at each CO.sub.2 level to assess the reproducibility of the signal changes associated with different CO.sub.2 levels. Each delivery of CO.sub.2 using Respiract was made in conjunction with arterial blood draw to determine the arterial blood CO.sub.2 levels. Imaging-based demonstration of myocardial hyperemic response to changes in PaCO.sub.2 was shown in health human volunteers with informed consent.
Example 1
[0153] We show that CO.sub.2 can increase myocardial perfusion by a similar amount, as does adenosine in canine models. We also show that in the setting of coronary artery narrowing, it is possible to detect regional variations in hyperemic response with the use of Mill by detecting signal changes in the myocardium due to changes in oxygen saturation (also known as the BOLD effect) using a free-breathing BOLD MRI approach.
[0154] As show in
[0155] In detail, the color images (lower panel of
Example 2
[0156] Co-Relation Between Inhaled CO.sub.2 and oxygen saturation
[0157] We assessed the difference between myocardial blood-oxygen-level dependent (BOLD) response under hypercarbia and normocarbia conditions in canines. The BOLD signal intensity is proportional to oxygen saturation.
[0158] Top panels of
[0159] We further assessed the myocardial BOLD response to stepwise CO.sub.2 increase (ramp-up) in canines. As shown in
[0160] To further evaluate vascular reactivity and coronary response to CO.sub.2, we measured the myocardial BOLD signal in response to block increases of CO.sub.2 in canines. Specifically, the myocardial BOLD signal was measured as the amount of CO.sub.2 administered to the canine subjects alternated between 40 mmHg CO.sub.2 and 50 mmHg CO.sub.2. As shown in
Example 3
[0161] Co-Relation Between the Amount of CO.sub.2 inhaled and Doppler flow
[0162] Doppler flow, an ultrasound-based approach which uses sound waves to measure blood flow, was used to show that administration of CO.sub.2 leads to vasodilation which results in greater blood flow, while PaO.sub.2 is held constant. The Doppler flow was measured in the left anterior descending (LAD) artery. As shown in
Example 4
[0163] Each of the Arteries which Supply Blood to the Myocardium Responds to the CO.sub.2 levels
[0164] The myocardium is supplied with blood by the left anterior descending (LAD) artery, the right coronary artery (RCA) and the left circumflex (LCX) artery. We measured the blood flow through each of these arteries in response to increasing CO.sub.2 supply. As shown in
Example 5
[0165] Vascular Reactivity to CO.sub.2 comparable to adenosine
[0166] Vascular reactivity of three canines that were administered with adenosine was compared with the vascular reactivity of canines that were administered with CO.sub.2. As shown in
[0167] Further, as shown in
Example 6
[0168] To derive a theoretical understanding of how repeated measurements may affect the BOLD signal response, for a given BOLD response to PaCO.sub.2, Applicants performed numerical simulations of statistical fits assuming various peak hyperemic BOLD responses to two different PaCO.sub.2 levels (as in
Example 7
[0169] We investigated whether a physiologically tolerable hypercapnia stimulus (˜25-mmHg increase in PaCO.sub.2) can increase myocardial blood flow (MBF) to that observed with adenosine in three groups of canines: (i) without coronary stenosis; (ii) subjected to non-flow limiting coronary stenosis; and (iii) following pre-administration of caffeine. These studies were conducted by prospectively and independently controlling PaCO.sub.2 and combining it with .sup.13N-ammonia Positron Emission Tomography (PET) measurements, and the extent of effect on MBF due to hypercapnia was compared to adenosine.
[0170] The objectives of these studies were twofold: to investigate the effects of PaCO2 on MBF while minimizing contributions from factors that can unintentionally reduce or inaccurately report on sensitivity of PaCO2 on MBF; and to assess whether an independent, precise and rapid establishment of physiologically tolerable level of hypercapnia provides equivalent hyperemia as adenosine, a commonly used pharmacological stimulus for cardiac stress testing with and without pre-administration of caffeine. To address these aims, a clinically relevant animal model was used along with validated strategies for (i) precisely and rapidly establishing desired levels of PaCO.sub.2, while holding PaO.sub.2 constant; (ii) quantifying MBF in vivo; and (iii) image analysis to derive MBF values across the different coronary supply territories. We compared our findings to the effects of standard dose of adenosine in the same animal models with and without coronary stenosis to quantify MBF and flow deficit regions under peak tolerable PaCO2. To determine whether MBF response to PaCO.sub.2 overlaps the same mechanistic path as adenosine, we quantified MBF under hypercapnia and adenosine following caffeine administration. Studies were conducted as described in Yang, H-J et al., J. Nucl. Med. 58: 953-960 (2017), which is hereby incorporated by reference in its entirety.
[0171] Prospectively Targeted Hypercapnia as Potent Stimulator of MBF and its Use for Identifying Regional Impairments in MBF and MPR
[0172] We found that prospectively targeted hypercapnia was a potent stimulator of MBF. Data are given in
[0173]
[0174] Perfusion Defect Volumes and Visual Scoring Under Hypercapnia Versus Adenosine
[0175] Perfusion defect volumes and visual scoring under hypercapnia vs. adenosine were determined. The total reduction in perfusion volume (TRP, % LV) between stress and rest states is shown in
[0176] The results in
[0177] Effect of Preadministration of Caffeine on MBF Under Hypercapnia Versus Adenosine
[0178] Next the effect of pre-administration of caffeine on myocardial blood flow under hypercapnia vs. adenosine was determined. Mean global MBF and MPR following pre- and post-caffeine administration are shown in
[0179] In sum, these results show that in the absence of stenosis, mean MBF under hypercapnia was 2.1±0.9 ml/min/g and adenosine was 2.2±1.1 ml/min/g were significantly higher than at rest (0.9±0.5 ml/min/g, P<0.05); and were not different from each other (P=0.30). Under left-anterior descending coronary (LAD) stenosis, MBF increased in response to hypercapnia and adenosine (p<0.05, all territories) but the effect was significantly lower than in the LAD territory (with hypercapnia and adenosine; both p<0.05). Mean perfusion defect volumes measured with adenosine and hypercapnia were significantly correlated (R=0.85) and were not different (p=0.12).
[0180] Following pre-administration of caffeine, a known inhibitor of adenosine, resting MBF decreased and hypercapnia increased MBF but not adenosine (p<0.05). The results indicate that arterial blood CO.sub.2 tension when increased by 25 mmHg can induce MBF to the same level as a standard dose of adenosine.
[0181] The results thus demonstrate substantial similarity in physiologic outcomes on several scores between a clinically-relevant hyperemia-inducing drug (adenosine) and carbon dioxide. It is noted that, in addition to having significant negative side effects, adenosine's negative effects can last for hours whereas carbon dioxide can be exhaled off in a few breaths. Importantly, it is not only the hyperemia inducing effect on the heart as a whole that is comparable for carbon dioxide and adenosine; the identified blood flow deficit is also the same (for both location and relative volume of deficit, which influences treatment choices in the clinic). There was also excellent concordance in how blind observers (blind to whether adenosine or CO.sub.2 was used to cause the effect) scored the images in terms of deficit severity. Thus, CO.sub.2 is able to discriminate ischemic heart disease in a clinically relevant setting, indicating that prospectively targeted arterial CO.sub.2 can be used as an alternative to current pharmacological vasodilators for cardiac stress testing.
[0182] Various embodiments of the invention are described above in the Detailed Description. While these descriptions directly describe the above embodiments, it is understood that those skilled in the art may conceive modifications and/or variations to the specific embodiments shown and described herein. Any such modifications or variations that fall within the purview of this description are intended to be included therein as well. Unless specifically noted, it is the intention of the inventors that the words and phrases in the specification and claims be given the ordinary and accustomed meanings to those of ordinary skill in the applicable art(s).
[0183] The foregoing description of various embodiments of the invention known to the applicant at this time of filing the application has been presented and is intended for the purposes of illustration and description. The present description is not intended to be exhaustive nor limit the invention to the precise form disclosed and many modifications and variations are possible in the light of the above teachings. The embodiments described serve to explain the principles of the invention and its practical application and to enable others skilled in the art to utilize the invention in various embodiments and with various modifications as are suited to the particular use contemplated. Therefore, it is intended that the invention not be limited to the particular embodiments disclosed for carrying out the invention.
[0184] While particular embodiments of the present invention have been shown and described, it will be obvious to those skilled in the art that, based upon the teachings herein, changes and modifications may be made without departing from this invention and its broader aspects. It will be understood by those within the art that, in general, terms used herein are generally intended as “open” terms (e.g., the term “including” should be interpreted as “including but not limited to,” the term “having” should be interpreted as “having at least,” the term “includes” should be interpreted as “includes but is not limited to,” etc.).