FIGURE SUMMARY
Title

Platinum-triggered Bond-cleavage of Pentynoyl amide and N-propargyl handles for Drug-Activation

Authors
Oliveira, B.L., Stenton, B.J., V B, U., de Almeida, C.R., Conde, J., Negrão, M., Schneider, F.S.S., Cordeiro, C., Godinho Ferreira, M., Caramori, G.F., Domingos, J.B., Fior, R., Bernardes, G.J.L.
Source
Full text @ J. Am. Chem. Soc.

CisPt Decages the Fluorogenic Probe 9 in vivo. Zebrafish larvae were exposed to 9 diluted in embryonic medium for 24 h, followed by a 1 h wash in embryonic medium. Larvae were randomly distributed into two conditions: DMSO or CisPt for 24 h (a). Confocal image of zebrafish larvae exposed to 9 + DMSO (b) and 9 + CisPt (c).

CisPt-mediated prodrug decaging in zebrafish xenografts. HCT116 human CRC cells were fluorescently labeled with lipophilic CM-DiI (shown in red) and injected into the perivitelline space (PVS) 2 days post fertilization (dpf) Tg(Fli1:eGFP) zebrafish larvae. Zebrafish xenografts were randomly distributed into treatment groups, daily treated with DMSO, CisPt, pFU, and pFU+CisPt and analyzed at 4, 6, or 7dpi for proliferation, apoptosis and tumor size. At 4 dpi, 6 dpi, and 7dpi, zebrafish xenografts were imaged by stereoscope (a–l) and by confocal microscopy (a′–lDAPI plus DiI, a′′–l′′ maximum projection of activated caspase 3). Proliferation (mitotic figures: mp, *P = 0.0104, ***P = 0.0004, ****P < 0.0001; s, **P = 0.0023, *** P = 0.0002), apoptosis (activated caspase 3: n, **P = 0.0033, ***P = 0.0006; q, *P = 0.0126, ****P < 0.0001; t, **P = 0.0068) and tumor size (n° of tumor cells: o, *P = 0.0279; r, ****P < 0.0001; u, *P = 0.0411, **P = 0.0010) were analyzed and quantified. Graphs represent fold induction (normalized values to controls) of Avg ± SEM. The number of xenografts analyzed is indicated in the representative images and each dot represents one zebrafish xenograft. Statistical analysis was performed using an unpaired test. Statistical results: ns > 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, and ****P ≤ 0.0001. All images are anterior to the left, posterior to right, dorsal up, and ventral down. Scale bar 50 μm.

Cisplatin-mediated prodrug decaging in a colon cancer Zebrafish Xenograft model at 6dpi. HCT116 human CRC cells were fluorescently labelled with lipophilic dye CM-DiI (shown in red) and injected into the PVS of 2 dpf Tg(Fli1:eGFP) zebrafish larvae. Zebrafish xenografts were treated in vivo for 5 consecutive days with: DMSO, 5-FU, CisPt, cFU, pFU, cFU+CispPt, pFU+CisPt and 5-FU+CisPt. At 6dpi (5dpt), zebrafish xenografts were imaged by stereoscope (A-H) and by confocal microscopy (A’-H’ DAPI plus DiI; A’’-H’’ a maximum projection of activated caspase 3). Quantification of proliferation (mitotic figures, I *** P=0.0004, ** P=0.0012, **** P<0.0001, * P=0.0106, * pFU vs pFU+CisPt P=0.0104, * cFU+CisPt vs pFU+CisPt P=0.0470), apoptosis (activated caspase3, J ****P<0.0001, * P=0.0126, ** P=0.0084, *** P=0.0002) and tumor size (no of tumor cells, K *** P=0.0008, * cFU P=0.0216, **** P<0.0001, * 5-FU+CisPt P=0.0273, ** P=0.0017). Outcomes are expressed by fold induction (normalized values to controls) as AVG ± SEM. The number of xenografts analyzed is indicated in the representative images and each dot represents one zebrafish xenograft. Statistical analysis was performed using an unpaired t-test. Statistical results: ns>0.05, *P≤0.05, **P≤0.01, ***P≤0.001 and ****P≤0.0001. All images are anterior to the left, posterior to right, dorsal up, and ventral down. Scale bar 50 μm.

Cisplatin-mediated prodrug decaging in a colon cancer Zebrafish Xenograft model at 7dpi. HCT116 human CRC cells were fluorescently labelled with lipophilic dye CM-DiI and injected into the PVS of 2 dpf Tg(Fli1:eGFP) zebrafish larvae. Zebrafish xenografts were treated in vivo for 6 consecutive days with: DMSO, Cispt, cFU, pFU, cFU+CisPt and pFU+CisPt. At 7dpi (6dpt), zebrafish xenografts were imaged by stereoscope (A-F) and by confocal microscopy (A’-F’ DAPI plus DiI, A’’-F’’ a maximum projection of activated caspase 3). Quantification of proliferation (mitotic figures, G * P=0.0180, ** P=0.0023, *** P=0.0002), apoptosis (activated caspase3, H * P=0.0119, ** P=0.0068) and tumor size (no of tumor cells, I ** P=0.0010, * cFU+CisPt vs pFU+CisPt P=0.0186, * pFU vs pFU+CisPt P=0.0411). Outcomes are expressed by fold induction (normalized values to controls) as AVG ± SEM. The number of xenografts analyzed is indicated in the representative images and each dot represents one zebrafish xenograft. Statistical analysis was performed using an unpaired t-test. Statistical results: ns>0.05, *P≤0.05, **P≤0.01, ***P≤0.001 and ****P≤0.0001. All images are anterior to the left, posterior to right, dorsal up, and ventral down. Scale bar 50 μm.


Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ J. Am. Chem. Soc.