Classification of prostate cancer using a protease activity nanosensor library

Abstract

Improved biomarkers are needed for prostate cancer, as the current gold standards have poor predictive value. Tests for circulating prostate-specific antigen (PSA) levels are susceptible to various noncancer comorbidities in the prostate and do not provide prognostic information, whereas physical biopsies are invasive, must be performed repeatedly, and only sample a fraction of the prostate. Injectable biosensors may provide a new paradigm for prostate cancer biomarkers by querying the status of the prostate via a noninvasive readout. Proteases are an important class of enzymes that play a role in every hallmark of cancer; their activities could be leveraged as biomarkers. We identified a panel of prostate cancer proteases through transcriptomic and proteomic analysis. Using this panel, we developed a nanosensor library that measures protease activity in vitro using fluorescence and in vivo using urinary readouts. In xenograft mouse models, we applied this nanosensor library to classify aggressive prostate cancer and to select predictive substrates. Last, we coformulated a subset of nanosensors with integrin-targeting ligands to increase sensitivity. These targeted nanosensors robustly classified prostate cancer aggressiveness and outperformed PSA. This activity-based nanosensor library could be useful throughout clinical management of prostate cancer, with both diagnostic and prognostic utility.

Theranostic Layer‐by‐Layer Nanoparticles for Simultaneous Tumor Detection and Gene Silencing

“…To confirm that diagnostic utility was preserved, LbL NPs were modified with biosensor and PEG or biosensor, iRGD, and PEG to yield NP+SP and NP+SRP, respectively. These NPs were then exposed to recombinant MMP9 (12.5 n m ) to verify protease‐induced cleavage of the reporter fragment from the NP . The MMP9‐digested NPs were pelleted by centrifugation and the supernatant was analyzed for the peptide reporter fragment (PRF) using an enzyme‐linked immunosorbent assay (ELISA) (Figure c).…”

Tissue-ABPP enables high-resolution confocal fluorescence imaging of serine hydrolase activity in cryosections – Application to glioma brain unveils activity hotspots originating from tumor-associated neutrophils

“…We applied tissue-ABPP also successfully to spleen sections, mainly to provide additional validation in native immune tissue for TAMRA-FP labeling in relation to some of the antibodies (Figures S34 and S35). Previously, a single study has presented low-resolution TAMRA-FP imaging of AEBSF-sensitive SH activity in prostate cancer specimen in supplementary figure (Dudani et al, 2018). Although preliminary, that work extends the applicability of tissue-ABPP to human proteomes.…”

Theranostic Layer‐by‐Layer Nanoparticles for Simultaneous Tumor Detection and Gene Silencing

“…To confirm that diagnostic utility was preserved, LbL NPs were modified with biosensor and PEG or biosensor,i RGD, and PEG to yield NP + SP and NP + SRP,respectively.These NPs were then exposed to recombinant MMP9 (12.5 nm)t o verify protease-induced cleavage of the reporter fragment from the NP. [19] TheM MP9-digested NPs were pelleted by centrifugation and the supernatant was analyzed for the peptide reporter fragment (PRF) using an enzyme-linked immunosorbent assay (ELISA) (Figure 1c). Ther esults indicated that the biosensor on the surface of the LbL NP is still accessible to the MMP9 protein, providing detectable peptide signal in the supernatant for both NP + Sa nd NP + SR. After initial validation of stability and function, the diagnostic capabilities of the biosensor LbL NPs were assessed across mouse models of pancreatic (subcutaneous flank tumor), colorectal (metastatic model), and ovarian cancer (flank and metastatic models).…”