Abstract
Biomarkers of disease progression and treatment response are urgently needed for patients with lymphangioleiomyomatosis (LAM). Activity-based nanosensors, an emerging biosensor class, detect dysregulated proteases in vivo and release a reporter to provide a urinary readout of disease. Because proteases are dysregulated in LAM and may directly contribute to lung function decline, activity-based nanosensors may enable quantitative, real-time monitoring of LAM progression and treatment response. We aimed to assess the diagnostic utility of activity-based nanosensors in a preclinical model of pulmonary LAM.
Tsc2-null cells were injected intravenously into female nude mice to establish a mouse model of pulmonary LAM. A library of 14 activity-based nanosensors, designed to detect proteases across multiple catalytic classes, was administered into the lungs of LAM mice and healthy controls, urine was collected, and mass spectrometry was performed to measure nanosensor cleavage products. Mice were then treated with rapamycin and monitored with activity-based nanosensors. Machine learning was performed to distinguish diseased from healthy and treated from untreated mice.
Multiple activity-based nanosensors [PP03 (cleaved by metallo, aspartic, and cysteine proteases), padj<0.0001; PP10 (cleaved by serine, aspartic, and cysteine proteases), padj=0.017)] were differentially cleaved in diseased and healthy lungs, enabling strong classification with a machine learning model (AUC=0.95 from healthy). Within two days after rapamycin initiation, we observed normalisation of PP03 and PP10 cleavage, and machine learning enabled accurate classification of treatment response (AUC=0.94 from untreated).
Activity-based nanosensors enable noninvasive, real-time monitoring of disease burden and treatment response in a preclinical model of LAM.
Footnotes
This manuscript has recently been accepted for publication in the European Respiratory Journal. It is published here in its accepted form prior to copyediting and typesetting by our production team. After these production processes are complete and the authors have approved the resulting proofs, the article will move to the latest issue of the ERJ online. Please open or download the PDF to view this article.
Conflict of interest: Dr. Kirkpatrick reports other from National Institute of Environmental Health Sciences, grants from Ludwig Fund for Cancer Research, grants from Koch Institute Marble Center for Cancer Nanomedicine, during the conduct of the study; In addition, Dr. Kirkpatrick has a patent Bhatia SN, Kirkpatrick JD, Dudani JS, Buss C, Warren AD. Lung protease nanosensors and uses thereof. International Patent Application PCT/US2019/052868 filed Sept. 25, 2019 pending.
Conflict of interest: Dr. Soleimany has nothing to disclose.
Conflict of interest: Dr. Dudani has nothing to disclose.
Conflict of interest: Dr. Liu has nothing to disclose.
Conflict of interest: Dr. Lam has nothing to disclose.
Conflict of interest: Dr. Priolo has nothing to disclose.
Conflict of interest: Dr. Henske has nothing to disclose.
Conflict of interest: Dr. Bhatia reports other from Howard Hughes Medical Institute, other from National Institute of Environmental Health Sciences, grants from Ludwig Fund for Cancer Research, grants from Koch Institute Marble Center for Cancer Nanomedicine, during the conduct of the study; other from Vertex Pharmaceuticals, other from Glympse Bio, other from Maverick Therapeutics, other from Satellite Bio, other from CEND Rx, other from Moderna Therapeutics, outside the submitted work; In addition, Dr. Bhatia has a patent Bhatia SN, Kirkpatrick JD, Dudani JS, Buss C, Warren AD. Lung protease nanosensors and uses thereof. International Patent Application PCT/US2019/052868 filed Sept. 25, 2019 pending.
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- Received March 4, 2021.
- Accepted August 14, 2021.
- Copyright ©The authors 2021. For reproduction rights and permissions contact permissions{at}ersnet.org
