Funding Distribution

Researchgrants54.3%

Contracts45.7%

Othercontracts

21.0%

NIHcontracts

24.7%

Funding Distribution

The MRL receives funding from government and non-governmental sources.

Funding Sources

Non-profitorganizations

24.3%

Privatesponsors

2.3%

Other federalsources

0.2%

National Institutesof Health

73.2%

Funding Sources

Innovations from the MRL have led to numerous patents and licensing agreements.

2014-2015

Licenses

Patent applications

Invention disclosures

0 5 10 15 20 25 30

Methodologies and approaches employed and developed at the MRL include molecular genetics, carbohydrate chemistry, protein chemistry, lipid chemistry, synthetic chemistry, enzymology, immunology, immunopathology, animal models of infection, and “omics’ technologies (genomics, metabolomics, proteomics, and lipidomics).

Recent MRL Contributions

Tuberculosis (TB)• Disease characterization

Diabetes is a risk factor linked to increased susceptibility to M. tuberculosis. MRL researchers have developed a guinea pig model of type 2 diabetes-TB comorbidity that mimics what is observed in humans with both diseases. In diabetic animals, TB infections were more severe and progressed more rapidly, and were associated with increased inflammation and a stronger proinflammatory response. This new model could lead to better understanding of the interplay of the two diseases, and to improved management of diabetic patients who acquire TB.[7]

• Therapeutics

MRL researchers and collaborators identified a compound effective at killing M. tuberculosis and having a mode of action different from currently used anti-TB treatments. The team discovered that the compound works by blocking MmpL3, a previously unidentified transporter of mycobacterial cell wall components that is required for mycobacterial growth.[8]

MRL scientists were part of an international team that identified a new candidate treatment for TB. The compound, called Q203, was highly effective against M. tuberculosis in culture and in a mouse TB model. The potency and pharmacokinetic and safety profiles of Q203 suggest it is an excellent candidate for clinical trials. Additionally, characterizing how Q203 kills TB reveals a new potential target for the development of future anti-TB therapeutics.[9]

• Vaccines

A team from the MRL demonstrated that the current approach to evaluating TB vaccines is flawed. The researchers used both mouse and guinea pig models to examine the efficacy of a TB vaccine and found substantial differences in the effectiveness of the vaccine against different TB strains. The vaccine was extremely effective against strains isolated in a region of South Africa where a recent trial of a new vaccine failed to show improvement compared to the existing vaccine. Based on their results, the MRL researchers suggest that the South African trial failed to show improvement because the TB strains circulating in the population in which the trial was carried out are so susceptible to the standard vaccine that no improvement is possible. The authors suggest a paradigm shift in the approach to TB vaccine trials, in which vaccines are first tested in animal models against the prevailing local bacterial strains before clinical field trials are conducted.[10]

• Surveillance/Diagnostics

Sputum smear microscopy is generally used to diagnose TB. Dr. Lenaerts of the MRL, in collaboration with colleagues from CSU and Massachusetts, described a new fluorescent stain able to detect 99% of M. tuberculosis in culture, a substantial improvement over traditional staining methods. The new method could improve detection of the bacteria in sputum from potential TB cases; almost half of such cases are missed using current methods.[11]

A less time consuming and more sensitive alternative to sputum smear microscopy would aid in TB diagnosis. MRL scientists, in collaboration with researchers in Switzerland, developed a sensitive assay using gas chromatography/mass spectrometry to detect and quantify lipoglycan lipoarabinomannan (LAM) in urine. The team demonstrated that LAM in urine is a powerful biomarker for active TB.[12]

Leprosy• Disease characterization

MRL researchers are examining how new cases of leprosy occur in individuals who have no known exposure to other people with the disease. Dr. Jackson and colleagues found that M. leprae could be taken up by free-living amoebae and remain viable and retain the ability to infect mice after spending up to 8 months within amoebic cysts. These results suggest one potential avenue through which M. leprae may survive in the environment between hosts.[13]

• Surveillance/Diagnostics

As cases of leprosy decrease, the expertise required to diagnose leprosy is also decreasing, highlighting a need for a simple diagnostic test for the disease.[14] Using a metabolomics approach, researchers at the MRL have identified differences between the serum of uninfected individuals and those recently infected with leprosy. Characterizing these differences may provide the first step towards identifying biomarkers to help diagnose or monitor the progression of leprosy.[15]

Non-tuberculous mycobacteria (NTM)• Disease characterization

Worldwide, patients with cystic fibrosis (CF) have been observed to acquire pulmonary infections of the NTM M. abscessus. As part of an international team, MRL scientists participated in a study to sequence the DNA of M. abscessus bacteria isolated from infected CF patients around the world. The study demonstrated that many recent infections are not independently acquired from the environment as previously thought, but instead appear to be acquired by indirect transmission within hospitals. The results have important implications for patient management. Interestingly, the same dominant strains are circulating around the world but the means of global spread are not known.[16]

• Therapeutics

Several ongoing drug discovery programs in the MRL labs aim to develop new compounds active against slow- and fast-growing NTM.

• Vaccines

MRL labs are optimizing animal models for NTM vaccine testing and developing candidate vaccines against NTM diseases.

Buruli ulcer• Disease characterization

MRL researchers reported changes in the lipid metabolic pathways of mosquitos contaminated with live M. ulcerans. The data presented indicate that mosquitos may help M. ulcerans survive and persist in aquatic environments in endemic areas. The study provides the basis for future research that may lead to strategies for intervening in this process.[17]

Mycobacterial infections of domestic and wild animals• Disease characterization

Dr. Olea-Popelka and international colleagues have recently highlighted the need to obtain a better understanding of zoonotic TB caused by M. bovis. The authors identify several immediate needs, including improved surveillance to achieve an accurate assessment of the incidence of zoonotic TB, improved diagnosis of zoonotic TB since it can present differently from TB and not be recognized by physicians, and studies to determine whether zoonotic TB, considered primarily a foodborne disease, can also be transmitted between people through the air. Also, because M. bovis is resistant to one of the first-line drugs used to treat TB and M. bovis infections require longer courses of therapy, identification of the underlying infectious agent is important to implementing appropriate treatment for zoonotic TB.[18-20]

Diseases studied at the MRL include:• Tuberculosis (TB) is curable, yet remains the most deadly infectious disease and one of the 10 leading causes

of death in the world.[1] Every year, more than 10 million new TB cases occur and 1.8 million people will die of the disease, 95% of whom live in low- and middle-income countries and 22% of whom are also infected with HIV.[1] Antibiotic resistance is of increasing concern, with 480,000 new antibiotic-resistant cases identified per year.[1]

• Leprosy, also known as Hansen’s disease, is a chronic disease affecting the skin, peripheral nerves, respiratory tract, and eyes. Leprosy is curable and international efforts incorporating the provision of cost-free multi-drug treatment have successfully eradicated leprosy from most of the Earth. However, with an incubation period that averages 5 years and can be as long as 20 years, leprosy can linger in populations and pockets of disease remain. Hundreds of thousands of people a year continue to become infected.[2]

• Buruli ulcer is a chronic, debilitating disease that primarily affects children.[3] Buruli ulcer leads to permanent disfigurement and disability but can be cured with eight weeks of antibiotics. Early detection and treatment are key to preventing or minimizing the effects of the disease. The infection is acquired from the environment but how remains unknown. A better understanding of how children acquire the disease is needed.

• More than 250 species of non-tuberculous mycobacteria (NTM) have been identified. Although primarily considered opportunistic pathogens, some may cause pulmonary, skin/soft tissue, lymphatic, central nervous system, and disseminated infections in individuals with predisposing conditions. Alarmingly, some of the most pathogenic NTM are a leading cause of pulmonary infections in patients with cystic fibrosis and the prevalence of infections caused by these NTM is increasing globally,[4] with very few treatment options available.[5]

• Animal and zoonotic mycobacterial infections of domestic and wild animals such as TB and Johne’s disease represent a substantial economic impact for agriculture and a public health concern due to zoonotic infections.[6]

In addition to grant-funded research, the MRL contracts with multiple partners to evaluate candidate therapeutics and vaccines, as well as provide specialized mycobacterial research reagents. The MRL is a major provider of preclinical services for the mycobacterial community. In 2014-2015, the MRL tested over 100 candidate antimycobacterial drugs and 50 vaccines under different formulations and in different animal models. Through dedicated NIH IDIQ contracts, the MRL prepared more than 5,000 units of TB and leprosy materials and reagents that were distributed through BEI Resources to 34 different countries, including 12 countries most affected by the burden of TB and leprosy.

32 4

WORK WITH THE MRLThe MRL continues to pursue excellence in research and training and welcomes new opportunities for collaboration and to leverage the expertise of MRL scientists in TB and leprosy research to address emerging mycobacterial pathogens.

To learn more about MRL faculty and their research interests, visit our website at http://mrl.colostate.edu/.

To learn more about contract services or to obtain specialized research reagents visit http://mrl.colostate.edu/services-reagents/.

To support the MRL’s educational and outreach activities visit https://advancing.colostate.edu/MRL.

COLORADO STATE UNIVERSITYCSU is a leading research university (Carnegie Classification R1) located in Fort Collins, Colorado. The university receives more than 300 million dollars a year in research funding. Its 27,000 undergraduate and 4,500 graduate and professional students are enrolled in 190 programs of study across eight colleges. The CVMBS at CSU grants more than 300 graduate degrees per year, including 130 D.V.M. degrees from its veterinary program, which is consistently rated as one of the top three schools in the nation.

REFERENCES1. Tuberculosis Fact Sheet. World Health Organization. October 2016; Available from: http://www.who.int/mediacentre/factsheets/

fs104/en/.

2. Leprosy Fact Sheet. World Health Organization. February 2017; Available from: http://www.who.int/mediacentre/factsheets/fs101/en/.

3. Buruli ulcer (Mycobacterium ulcerans infection). World Health Organization. February 2017; Available from: http://www.who.int/mediacentre/factsheets/fs199/en/.

4. Johnson, M.M. and Odell, J.A., Nontuberculous mycobacterial pulmonary infections. J Thorac Dis, 2014. 6(3): p. 210-20.

5. Ryu, Y.J., Koh W.J., and Daley, C.L., Diagnosis and treatment of nontuberculous mycobacterial lung disease: clinicians’ perspectives. Tuberc Respir Dis, 2016. 79(2): p. 74-84.

6. Palmer, M.V., et al., Mycobacterium bovis: a model pathogen at the interface of livestock, wildlife, and humans. Vet Med Int, 2012. 2012: p. 236205.

7. Podell, B.K., et al., Increased severity of tuberculosis in Guinea pigs with type 2 diabetes: a model of diabetes-tuberculosis comorbidity. Am J Pathol, 2014. 184(4): p. 1104-18.

8. Grzegorzewicz, A.E., et al., Inhibition of mycolic acid transport across the Mycobacterium tuberculosis plasma membrane. Nat Chem Biol, 2012. 8(4): p. 334-41.

9. Pethe, K., et al., Discovery of Q203, a potent clinical candidate for the treatment of tuberculosis. Nat Med, 2013. 19(9): p. 1157-60.

10. Henao-Tamayo, M., et al., The efficacy of the BCG vaccine against newly emerging clinical strains of Mycobacterium tuberculosis. PLoS One, 2015. 10(9): p. e0136500.

11. Ryan, G.J., Shapiro, H.M., and Lenaerts, A.J., Improving acid-fast fluorescent staining for the detection of mycobacteria using a new nucleic acid staining approach. Tuberculosis, 2014. 94(5): p. 511-8.

12. De, P., et al., Estimation of D-Arabinose by gas chromatography/mass spectrometry as surrogate for mycobacterial lipoarabinomannan in human urine. PLoS One, 2015. 10(12): p. e0144088.

13. Wheat, W.H., et al., Long-term survival and virulence of Mycobacterium leprae in amoebal cysts. PLoS Negl Trop Dis, 2014. 8(12): p. e3405.

14. Roset Bahmanyar, E., et al., Leprosy diagnostic test development as a prerequisite towards elimination: requirements from the user’s perspective. PLoS Negl Trop Dis, 2016. 10(2): p. e0004331.

15. Al-Mubarak, R., et al., Serum metabolomics reveals higher levels of polyunsaturated fatty acids in lepromatous leprosy: potential markers for susceptibility and pathogenesis. PLoS Negl Trop Dis, 2011. 5(9): p. e1303.

16. Bryant, J.M., et al., Emergence and spread of a human-transmissible multidrug-resistant nontuberculous mycobacterium. Science, 2016. 354(6313): p. 751-757.

17. Hoxmeier, J.C., et al., Analysis of the metabolome of Anopheles gambiae mosquito after exposure to Mycobacterium ulcerans. Sci Rep, 2015. 5: p. 9242.

18. Olea-Popelka, F., et al., Zoonotic tuberculosis in human beings caused by Mycobacterium bovis-a call for action. Lancet Infect Dis, 2017. 17(1): p. e21-e25.

19. Fujiwara, P.I. and Olea-Popelka, F., Editorial commentary: why it is important to distinguish Mycobacterium bovis as a causal agent of human tuberculosis. Clin Infect Dis, 2016. 63(5): p. 602-3.

20. Miller, M. and Olea-Popelka, F., One Health in the shrinking world: experiences with tuberculosis at the human-livestock-wildlife interface. Comp Immunol Microbiol Infect Dis, 2013. 36(3): p. 263-8.

21. World TB Day 2017. Center for Disease Control. Available from: https://www.cdc.gov/tb/worldtbday/.

M Y C O B A C T E R I AR E S E A R C H L A B O R A T O R I E SEmphasis on the Complex Biology of Pathogens and Hosts

Antibiotic Resistance and Mode of Action

Discovery and Testing of New Drugs and Diagnostics

Vaccine Development

Generation of Novel Models of Disease

A UNIQUE ENVIRONMENT FOR COLLABORATION AND TRANSLATIONAL RESEARCHThe MRL is situated within a unique environment with access to exceptional resources and facilities. The MRL is one of several research programs within CSU’s Department of Microbiology, Immunology and Pathology (DMIP) in the College of Veterinary Medicine and Biomedical Sciences (CVMBS). The MRL is highly integrated and collaborates extensively internally as well as externally. MRL partners with a number of private companies, non-profit foundations, and research and clinical laboratories around the world, including field studies in endemic areas.

Through the Infectious Disease Research Center (IDRC), MRL scientists have access to safe, secure, and state-of-the-art biosafety research and manufacturing facilities, including biosafety level 3 (BSL-3) bacteriology, immunology, and animal facilities. Multiple animal models of mycobacterial diseases have been developed by MRL scientists. The MRL is NIH/NIAID partner offering services including drug screening and vaccine efficacy evaluation. Additionally, the MRL is a worldwide source of specialized TB and leprosy research materials and reagents through the Biodefense and Emerging Infections Research Resources Repository.

Specialized facilities Preclinical servicesMycobacterial

research reagents• 20,000 sq. ft. of BSL-2 laboratories

• 10,000 sq. ft. of BSL-3 and ABSL-3 laboratories

• Histotechnology and Pathology Core

• Flow Cytometry Core

• Genomics and Proteomics Core

• Proteomics and Metabolomics Facility

• Central Instrumentation Facility (liquid and gas chromatography, NMR, mass spectrometry)

• Microscope Imaging Network

• Mycobacterial drug testing program

• Vaccine efficacy testing program

• Biodefense and Emerging Infections Research Resources Repository

Collaboration with infectious disease clinicians globally, the Colorado Clinical and Translational Sciences Institute (CCTSI), the Colorado Cystic Fibrosis Research and Development Program (National Jewish Health, Denver, CO), CSU Ventures, and BioMARC (BSL-3 and Select Agent cGMP Biopharmaceutical Manufacturing and Academic Resource Center at Colorado State University) provides tools and expertise to support translation of MRL discoveries to real world application and medical practice.

TRAINING AND EDUCATIONAL ACTIVITIESEducation and training of the next generation of scientists is an important mission of the MRL. The technologies employed and developed by MRL researchers for the study of mycobacteria are relevant to the fields of infectious diseases, antimicrobial resistance, microbiology, biochemistry, immunology, medicine, and diagnostic/biomarker/vaccine and drug discovery. At any one time, approximately 50 graduate and undergraduate students, postdoctoral fellows, and visiting scientists are housed within the MRL. MRL scientists contribute to the training of additional personnel from related departments through collaboration and coursework.

The MRL strives to disseminate information about mycobacterial diseases within the scientific community as well as to the general public. The second Colorado Mycobacteria Conference held in Fort Collins in 2016 attracted academic and clinical researchers, industry representatives, and nonprofit groups from around the world to share recent findings and foster collaboration. MRL researchers present dozens of invited talks every year, domestically and abroad.

Education is a key foundation of the worldwide effort to stop TB, a disease that is frequently misdiagnosed and undertreated in the United States.[21] To mark World TB Day 2016, MRL scientists hosted 80 students from local high schools who visited labs to learn about TB and gain hands-on laboratory experience. MRL researchers also hosted “Bacteria Wars” at the Fort Collins Museum of Discovery to teach museum goers about bacterial infections, including TB.

CCTSI

Arthropod-borneand Infectious Diseases

Laboratory (AIDL)

ESKAPE pathogens:biofilms and antimicrobial

resistance

MRL

Prion ResearchCenter (PRC)

Retroviral ResearchGroup (RRG)

TherapeuticsDiagnostics

VaccinesPreclinical Services

DMIPResearch Programs

Colorado Cystic FibrosisR&D Program

CSU Ventures

BioMARCThe MRL at Colorado State UniversityWHO WE AREThe Mycobacteria Research Laboratories (MRL) is the largest academic research group in the United States devoted to the study of mycobacteria and mycobacterial diseases such as tuberculosis (TB), leprosy, Buruli ulcer, and non-tuberculous mycobacterial infections. A world leader in basic, preclinical, and field research related to mycobacterial diseases, the MRL is situated within a unique, collaborative environment providing resources and expertise to speed translation of research findings to real-world clinical applications such as improved diagnostics, new therapies, and more effective vaccines. Through teaching, training, research, and outreach, MRL scientists have a global influence on the understanding and treatment of mycobacterial diseases.

A SUBSTANTIAL PUBLIC HEALTH PROBLEM ADDRESSED BY THE MRLTB is the most deadly infectious disease in the world. TB and other mycobacterial infections tend to be chronic and debilitating. Though many are curable, adherence to the long-term courses of treatment required for a cure can be difficult. New diagnostics to allow earlier and more specific diagnosis, new therapeutics to enable faster and easier treatment, and new or improved vaccines to prevent infection altogether could help eradicate these diseases, prevent millions of deaths, and reduce the human cost associated with disfigurement and disability. Despite the significant health and economic costs of mycobacterial diseases, large biopharmaceutical companies are largely absent from this space because the diseases disproportionately affect poorer countries. This makes the work and mission of groups such as the MRL all the more essential.

MRL Vision for the Control and Elimination of Mycobacterial Diseases

GoalsTherapeutics to Achieve Faster and Durable Cure

Vaccines to Prevent Disease and Transmission

Diagnostics that Contribute to the Control of Disease

Major Research Initiatives

Define the metabolism and regulation of Mycobacterium spp. in vivo

Develop animal models reflective of human disease

Develop diagnostic markers that enable use of point-of-care platforms

Elucidate drug exposure and synergy at the site of disease

Elucidate essential mediators of protective immunity

Develop biomarkers for prediction of disease progression and outcome

Discover combinations of effective host- and pathogen-directed therapies

Define environmental factors that influence the development and persistence of protective immunity

Identify companion diagnostics for vaccines and therapeutics

Elucidate biomarkers for early prediction of treatment outcome

Define the contribution of mycobacteria strain diversity on protective immunity

Develop diagnostics that allow for control of zoonotic transmission of disease

The MRL is at the forefront of research and training in mycobacterial disease, consistently producing high-impact work with real-word implications for understanding, diagnosing, treating, and preventing these diseases. Over the past six years, MRL scientists have published more than 420 publications and their work has been cited more than 4,000 times.

2014-2015

0

50

100

150

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250

POSTDOCTORAL FELLOWS

VISITING SCIENTISTS

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UNDERGRADUATE STUDENTS

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32165