Progress 09/15/22 to 08/29/24
Outputs
Target Audience:The target audience for the duration of the project were basic scientists, clinicians, and clinican-scientists in the fields of vascular medicine and vascular biology. Changes/Problems:I moved to the University of Kentucky from Baylor University, where I originally submitted the Postdoctoral Fellowship. However, I still accomplished the proposed goals, and moving also gaveme the opportunity to expand beyond them to also include work on the COCOA-PAD trial as well as to receive trianing in the gut microbiome field. What opportunities for training and professional development has the project provided?During thisproject, I have received training from my mentor, Dr. John McCarthy, on working with animal models. Prior to this project, I lacked experience using an animal model; thus, this training provided me with a unique skill set important for my goal of conducting translational research. My animal training included a full curriculum on animal care and use, and submission of my own Institurional Animal Care and Use Committee (IACUC) proposal. I also received one-on-one training on performing the hindlimb ischemia surgery described. I also was exposed to the field of the gut microbiome and received training on metagenomics, which led to an additional publication (PMID: 37940330) and one under review. Professional Development: I attended the American Heart Association Scientific Sessions 2022 (November 2022). I also attended the Northwestern University Walking and Circulation Study Investigator's Meeting (April 2024) as well as the American Aging Association Annual Meeting (June 2024). How have the results been disseminated to communities of interest?Data generated from this project have been published in peer-reviewed journals. The main publication was published inAJP-Cell Physiology, and the paper receieved theAPSselectAward. The paper was summarized on Newswise. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Impact Statement: This project led to a change in knowledge. We identified the molecular mechanisms underlying the therapeutic effect of natural food-derived antioxidants (epicatechin-rich cocoa and sulforaphane-rich broccoli) on wakling performance in peripheral artery disease (PAD). This work was published in?a peer-reviewed journal. Under the goal/objective of 1) Advancing science: This work aimed to give insights into the mechanisms of sulforaphane's actions and identify whether sulfroaphane may be a viable therapeutic approach in the context of peripheral artery disease (PAD), a condition which results in severe walking and functional limitations. To this end, we performed experiments in an animal model of PAD (hindlimb ischemia). Mice were fed either a control diet or a diet supplemented with sulforaphane-rich broccoli. We found that the mice fed broccoli-supplemented diets had improved clinical limb use scores at 6 and 12 weeks following the hindlimb ischemia surgery, indicating more normal walking compared to mice fed control diets. The mice fed broccoli-supplemented diets also had a reduction in skeletal muscle free radical production of almost 50%. We found that this was mediated by an activation of the Nrf2 protein in the muscle, which controls the production of several antioxidants. Skeletal muscle mitochondrial respiration was also improved in the broccoli-supplemented diet group. To further identify whether other natural compounds can increase Nrf2 activation in skeletal muscle of PAD patients, we obtained muscle biopsy samples from the COCOA-PAD Pilot Randomized Clinical Trial, which randomized participants with PAD to either a daily epicatechin-rich cocoa beverage or placebo for 6 months. We found that Nrf2 target antioxidants were signficantly increased in the skeletal muscle biopsies of participants from the cocoa group, but not in the placebo group. The increase in antioxidant protein levels was associated with reduced accumulation of central nuclei, a myopathy indicator, and increased protein levels of mitochondiral respiratory complex subunits. In vitro studies in primary human myotubes further showed that epicatechin and sulforaphane treatment protected against PAD serum-induced oxidative stress and mitochondrial decrements by preventing the reduction in Nrf2 activation. These data led to a change in knowledge, resulting in a peer-review manuscript publication. 2) Regarding the goal of informing future studies: The encouraging preclinical data will encourage a future clinical tiral to test sulforaphane-rich broccoli in PAD patients. Furthermore, the results from the pilot cocoa trial have led to a change in action, the modification of the ongoing COCOA-PAD II Trial to include assessment of Nrf2 measures. 3) The goal of the formal career development program will be outlined in the training and professional development below.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2024
Citation:
Ismaeel A, McDermott MM, Joshi JK, Sturgis JC, Zhang D, Ho KJ, Sufit R, Ferrucci L, Peterson CA, Kosmac K. Cocoa flavanols, Nrf2 activation, and oxidative stress in peripheral artery disease: mechanistic findings in muscle based on outcomes from a randomized trial. Am J Physiol Cell Physiol. 2024 Feb 1;326(2):C589-C605. doi: 10.1152/ajpcell.00573.2023. Epub 2024 Jan 8. PMID: 38189132; PMCID: PMC11193455.
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Koopmans PJ, Ismaeel A, Goljanek-Whysall K, Murach KA. The roles of miRNAs in adult skeletal muscle satellite cells. Free Radic Biol Med. 2023 Nov 20;209(Pt 2):228-238. doi: 10.1016/j.freeradbiomed.2023.10.403. Epub 2023 Oct 24. PMID: 37879420; PMCID: PMC10911817.
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Ismaeel A, Valentino TR, Burke B, Goh J, Saliu TP, Albathi F, Owen A, McCarthy JJ, Wen Y. Acetate and succinate benefit host muscle energetics as exercise-associated post-biotics. Physiol Rep. 2023 Nov;11(21):e15848. doi: 10.14814/phy2.15848. PMID: 37940330; PMCID: PMC10632089.
- Type:
Journal Articles
Status:
Under Review
Year Published:
2024
Citation:
Valentino TR, Burke BI, Kang G, Goh J, Dungan CM, Ismaeel A, Mobley CB, Franco AB, May JB, Flythe MD, Wen Y, McCarthy JJ. Microbial-Derived Exerkines Prevent Skeletal Muscle Atrophy. Under review in Nature Metabolism.
|
Progress 09/15/22 to 09/14/23
Outputs
Target Audience:The target audience during this reporting period were basic scientistis, clinicians, and clinician-scientists in the fields of vascular medicine and vascular biology. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Training Activities: During this reporting periodworking on this project, I have received training from my mentor, Dr. John McCarthy, on working with animal models. Prior to this project, I lacked experience using an animal model; thus, this training provided me with a unique skill set important for my goal of conducting translational research. My animal training included a full curriculum on animal care and use, and submission of my own Institurional Animal Care and Use Committee (IACUC) proposal. I also received one-on-one training on performing the hindlimb ischemia surgery described. Professional Development: I attended the American Heart Association Scientific Sessions 2022 (November 2022). How have the results been disseminated to communities of interest?The results have been submitted as a manuscript to a peer-reviewed scientific journal, which is currently under review. What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period, we will complete theproposed experiments. This includes increasing the sample size for the animal studies, and adding the proposed groups of mice fed a Western diet and a Western diet supplemented with broccoli. Further analysis of fixed muscle and artery sections will also be completed. Efforts will be made to ensure final acceptance and publication of the associated manuscript.
Impacts
What was accomplished under these goals?
IMPACT: Peripheral artery disease, or PAD, is caused by a blockage of the arteries that carry blood to the legs. PAD affects over 10 million people in the United States. Annual costs for PAD-related treatment in the U.S.are estimated to exceed $21 billion. PAD leads to severe functional limitations and reductions in quality of life of patients. There are very few treatment options for patients with PAD. Only two medications are approved by the FDA to treat walking impairments, but both medications show onlylimited benefit. One of the factors that leads to the reduced walking ability in patients with PAD is an increase in molecules called free radicals that can damage the muscles and arteries of the legs. One of the ways to reduce this damage isto increase the antioxidants that the body produces, which can counteract the free radicals. A protein called "Nrf2" controls the production of many antioxidants in the body. This protein can be activated by different compoundsfound in foods such as broccoli and cocoa. To test if we can use foods rich in these compounds to increase Nrf2 activity and improve outcomes in PAD, first I used an animal model of the disease. In this model,we perform a surgery where wetie off the leg arteries of mice, which causes similar symptoms as human PAD. One group of mice was fed a control diet, and another group of mice was fed a diet supplemented with broccoli. The mice fed broccoli-supplemented diets had improved clinical limb use scores at 6- and 12-weeks following the surgery. This means that they had more normal walking compared to the mice fed control diets without broccoli. The mice fed broccoli-supplemented diets also had almost 50% reduced free radical production in their muscles. Next, I obtained muscle biopsy samples from the COCOA-PAD Pilot Randomized Clinical Trial, which was a 6-month intervention that randomized participants with PAD to either a daily cocoa beverage or placebo. Patients receiving cocoa had significantly improved walking performance compared to the placebo group. In my studies, I looked at antioxidant proteins in the muscle biopsies and found that they were also increased in the group that received the cocoa beverage. Next, to further establish the mechanism, I cultured muscle cells and treated them with serum from the PAD patients from the COCOA-PAD Trial. The serum treatment increased free radicals and damage to the muscle cells. When I also treated cellswith the compounds found in broccoli or cocoa, these compounds reduced the free radicals and the damage caused to the muscle cells by the serum. These findings suggest that consumption of foods such as broccoli and cocoa that activate the Nrf2 pathway and increase antioxidant production may be promising therapeutic approaches for PAD treatment to reduce free radical production and improve walking performance. The ongoing COCOA-PAD II trial will confirm if this inexpensive, safe, accessible, and well-tolerated therapy has the potential to meaningfully improve mobility in people with PAD. The findings also encourage future studies in patients to see whether consuming broccoli can improve their walking performance. Goals and Objectives: 1) Advancing Science: This work will give insights into the mechanisms of sulforaphane's action. Peripheral artery disease is a growinghealth problem, and this project will identify whether sulforaphane may be a viable therapeutic approach in the context of limb ischemia, by targeting the pathophysiological mechanism of arterial dysfunction. The major activities completed/experiments conducted for this objective include the animal experiments discussed above. I have collected data showing that sulforaphane-rich broccoli-supplemented diets significantlyimprove walking ability in a mouse model of PAD and signficantly reduce free radical production. These represent a change in knowledge. These, along with the cocoa data presented above, have been written as a manuscript, which has been submitted to a peer-reviewed journal to be considered for publication. 2) This work will also inform future studies to test whether sulforaphane-containing cruciferous vegetables, such as broccoli/broccoli sprouts, may improve walking performance in peripheral artery disease patients. Successful publication of the preclinical data will encourage a clinical trial in patients testing this. 3) Additionally, the formal career development program outlined will allow the candidate to systematically develop skills necessary for PI development as an independent tenure-track investigator and teacher. This is described in the training and professional development below.
Publications
- Type:
Journal Articles
Status:
Under Review
Year Published:
2023
Citation:
Ismaeel A, McDermott MM, Joshi J, Sturgis JC, Zhang D, Ho K, Sufit RL, Ferrucci L, Peterson CA, Kosmac K. Cocoa Flavanols Blunt Peripheral Artery Disease-Associated Oxidative Stress via Activation of Nrf2. Submitted to Free Radical Biology and Medicine (Under Review).
|
|