Host biomarkers for investigating the spectrum of M. tuberculosis infection
| Background: | Evaluation of cytokines and chemokines that distinguish active TB, including extrapulmonary TB, from latent Mtb infection. Using archived, cryopreserved PBMC samples collected by Dr. Bruno Andrade in Salvador, Brazil, we are evaluating the performance of our candidate biomarkers in individuals with varying degrees of severity of Mtb infection and disease, including LTBI, sub-clinical TB and ATB; the latter will include both pulmonary and extrapulmonary disease. We will also assess our biomarkers in samples derived from individuals with presumptive TB (symptomatic or asymptomatic) for whom diagnostic tests were negative for TB. |
| Primary aims: | Evaluate the performance of T cell biomarkers for diagnosing TB across the spectrum of Mtb infection in HIV-uninfected and HIV-infected individuals. |
| Current status: | A paper describing our results was published in CID (please refer to Publications). |
| Principal Investigators (Consortium PIs): | Jyothi Rengarajan, Ph.D., Bruno Andrade, M.D., Ph.D. |
Prospective profiling of eicosanoid and inflammatory balance in TB-diabetes – CRDF Global
| Background: | This study utilize specimens only from Salvador site in Brazil and South Africa. Associations between the eicosanoid balance and TB clinical outcomes were further demonstrated in patients with pulmonary TB in an exploratory study from our group. A longitudinal multi-site study simultaneously quantifying several eicosanoids is needed to evaluate whether eicosanoid balance reflects TB disease activity and if/how this balance is influenced by HIV and/or DM. |
| Primary aims: | To define the clinical characteristics of individuals with TB and/or DM and/or HIV in Brazil and South Africa; To determine whether co-morbid DM is associated with qualitatively or quantitatively distinct eicosanoid profiles in patients with TB and/or TB-HIV; and to determine whether co-morbid DM is associated with qualitative or quantitative differences in the resolution of perturbed eicosanoid profile during and after ATT in patients with TB and/or TB-HIV. |
| Current status: | After obtaining the protocol approval by the IRBs from South Africa, Brazil, and Vanderbilt University Medical Center (VUMC), the study sites finalized patient recruitment. We also established a Materials Transfer Agreement (MTA) that allowed shipment of biospecimens from Brazil and South Africa to Vanderbilt. An export permit was also required to ship the specimens from South Africa to VUMC. In addition, the SOPs of the laboratory assays were also finalized. The lipidomic assays were completed, plasma specimens from cohort A (TB patients with or without diabetes) were successfully shipped from South Africa to Brazil for the Luminex assays. Results were presented during international meetings and papers were published. Additional manuscripts are also ongoing. Please refer to our Publications page. |
| Principal Investigators: | Bruno Andrade, M.D., Ph.D., Timothy Sterling, M.D., Henrique Serezani, Ph.D., John Koethe, M.D., Ginger Milne, M.S.C.I., Alisdair Leslie, Ph.D. |
Cross-RePORT TB-Diabetes transcriptomic signatures – CRDF Global
| Background: | Evaluation of transcriptomic signatures in persons with TB, TB/DM. This study was in collaboration with sites in India and South Africa. This study utilized specimens from the Salvador, Bahia site. |
| Primary aims: | This project used peripheral blood samples collected in RNA storage tubes by the RePORT studies in Brazil, South Africa and India. Samples were used for RNA sequencing to investigate the transcriptomic profile of TB patients with and without comorbid diabetes. |
| Current status: | RNA samples extracted from participants´ specimens in Brazil were sent to India for the transcriptomic analysis. Results from the Brazilian samples were generated together with South African and Indian samples. A manuscript is under review and accepted for publication. Please refer to our Publications page. |
| Principal Investigators: | Hardy Kornfeld, M.D., Bruno Andrade, M.D., Ph.D., Amita Gupta, M.D. |
Validation of transcriptional signature to predict active TB disease among advanced HIV patients – CRDF Global
| Background: | Evaluation of transcriptomic signatures in advanced HIV, and TB/HIV. This is in collaboration with RePORT-India. This study used specimens only from the INI and FMT Brazil sites. |
| Primary aims: | To conduct testing and validation of the 15-gene signature to predict active TB disease among advanced HIV patients with CD4 <100 cells/ul; and to assess and compare the cytokine/chemokine signature that may be predictive for active TB among advanced TB-HIV and HIV patients in India and Brazil. |
| Current status: | IRB approvals in Brazil were received and all patients were recruited. Samples were collected and stored. Luminex assays were complete and a paper was published. Additional manuscripts are planned. Please refer to our Publications page. |
| Principal Investigators: | Valeria Rolla, MD, PhD, Marcelo Cordeiro dos Santos, MD, PhD, Padmini Salgame, PhD, Vidya Mave, MD, MPH, Amita Gupta, MD, Bruno Andrade, MD, PhD. |
Mapping M. tuberculosis transmission hotspots using patient location history
| Background: | Mapping hotspots for transmission of infectious diseases can offer great help in controlling outbreaks and eliminating reservoirs of infection. Usually, hospital staff in endemic areas try to determine such places by asking patients with transmissible infections, those places where they live/work (for diseases which require chronic exposure) or visited a few days prior going to the hospital (for acute diseases). However, information obtained through questionnaires are generally vague, inaccurate and not integrated into databases. This makes the process manual, slow, and of little value for large-scale epidemiologic studies. Since a significant portion of the population has mobile phones with GPS, the objective of this project is to improve the accuracy and organization of M. tuberculosis infection networks, and to study the temporal dynamics of geolocalization data collected from TB patients. We developed an online tool in which patients who arrive at the referral centers can voluntarily provide GPS data of their mobile phones. User data are anonymized, processed and sent to a secure server. By analyzing the location patterns of hundreds of patients during several months prior the clinical visit, we will be able to potentially map hotspots of M. tuberculosis transmission. |
| Primary aims: | To use location history data from TB patients and their close contacts to find regions with a high risk of disease transmission. And to develop a web application to collect location history data from those patients diagnosed with TB and their close contacts, and implement a data analysis pipeline for finding the disease hotspots. |
| Current status: | IRB approvals in Brazil were received and patients recruited. Please refer to our Publications page. |
| Principal Investigators: | Helder Nakaya, PhD., Bruno Andrade, MD, PhD. |
TB/HIV: Predictors of treatment toxicity, failure, and relapse in HIV related TB – NIH
| Background: | This study evaluated pharmacogenomic predictors of TB/HIV treatment toxicity, and TB treatment outcomes (e.g., 2-month culture conversion and failure/relapse). All RePORT-Brazil study sites enrolled participants into this study and several manuscripts are published or under review (please refer to our Publications page). |
| Primary aims: | To identify pharmacogenomic predictors of TB/HIV plasma drug exposure and increased risk for toxicity during TB therapy; and to determine the pharmacogenomic predictors of two month culture-positivity, and TB treatment failure and relapse, while accounting for host and M. tuberculosis pathogen factors. |
| Current status: | Completed. Please refer to our Publications page. |
| Principal Investigators: | Timothy Sterling, M.D., Valeria Rolla MD, PhD., David Haas, M.D. |
Prospective, Multicentre Trial to Assess the Diagnostic Accuracy of the Truenat Assays at Intended Settings of Use – CNPq
| Background: | The Truenat MTB (including both MTB and MTB plus) and the MTB-RIF Dx reflex assays (Molbio Diagnostics; Bangalore, India) utilize chip-based real-time micro PCR for detection of tuberculosis (TB) and rifampicin (RIF) resistance from DNA extracted from sputum samples in about 25 minutes. A pilot trial conducted in India of the Truenat MTB assay found the assay to achieve high clinical performance. However, further evidence of the Truenat MTB as well as the Truenat MTB-RIF Dx assay, is needed prior to recommending the clinical use of the assays. |
| Primary aims: | To determine the diagnostic accuracy of the Truenat MTB assays and MTB-RIF Dx assay using culture and phenotypic/genotypic drug susceptibility test (DST) as gold standard in the intended setting of use. |
| Current status: | Completed. Please refer to our Publications page. |
| Principal Investigators: | Adam Penn-Nicholson (FIND, Switzerland), Afranio Kritski, M.D., Ph.D.; Sites in RePORT-India; Other sites in Peru, Ethiopia and Papua New Guinea. |
Evaluation of microvirin-based point-of-care diagnostic tests for tuberculosis
| Background: | Biomarker detection tests, such as enzyme-linked immunosorbent assays (ELISAs) and lateral flow assays (LFAs), are inexpensive and can be performed with minimal equipment. LFAs are particularly advantageous as they can provide results rapidly at the point-of-care (POC). The most commonly detected biomarker for TB is lipoarabinomannan (LAM). Currently, the only commercially-available LFA for LAM is the Alere LFA, which detects LAM in urine – a noninvasive, low biohazard risk sample method. However, due to low sensitivity, the Alere LFA is only approved for use in HIV-positive individuals as immunocompromised individuals have higher concentrations of LAM in urine. With increased sensitivity, an LFA could detect LAM in all individuals, regardless of immune status. Such an LFA would be an important advance in TB diagnostic tests, given its rapid turn-around-time and low-cost. Microvirin (MVN) is a lectin with a high affinity for alpha-1,2-mannose linkages, which are present in the endcaps of M.tb LAM. Using bio-layer interferometry, we have studied the binding of MVN to LAM and found the interaction to be of equivalent strength or stronger than the binding of LAM to anti-LAM antibodies. Furthermore, MVN has enhanced specificity for M. tb. Anti-LAM antibodies bind to the LAM backbone, which is common to LAM from all species of mycobacteria. In contrast, MVN binds to the endcaps of LAM, which vary according to the species of Mycobacterium. Thus, using MVN as a molecular recognition element to detect LAM in an LFA for diagnosis of TB would potentially result in a more sensitive and specific test compared to traditional LAM biomarker detection tests that utilize antibodies, while maintaining the speed and low cost that make LFAs so promising. |
| Primary aims: | The purpose of this study is to evaluate the clinical performance of two diagnostic tests for TB: an on-bead ELISA and a lateral flow assay. Both tests use MVN to capture and detect LAM, a biomarker for TB, in urine. |
| Current status: | Completed. Please refer to our Publications page. |
| Principal Investigators: | David Wright, Ph.D., Megan van der Horst, Micaella Jorge, Valeria Rolla, M.D., Ph.D, Adriano Gomes, Ph.D, Timothy Sterling, M.D. |