A topically-sprayable, activatable fluorescent and retaining probe, SPiDER-βGal for detecting cancer: Advantages of anchoring to cellular proteins after activation

Nakamura, Yuko; Mochida, Ai; Nagaya, Tadanobu; Okuyama, Shuhei; Ogata, Fusa; Choyke, Peter L.; Kobayashi, Hisataka

doi:10.18632/oncotarget.17080

Cited by 20 publications

(25 citation statements)

References 32 publications

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“…Skin is used extensively in many areas of research such as analysing the effects of irradiation (Kim et al, ), studying the absorption and effects of transdermal drugs (Mendoza‐Garcia, Sebastian, Alonso‐Rasgado, & Bayat, ), screening toxicity of topically applied compounds (Nakamura et al, ) and studying wound repair (Leal, Cordery, Delgado‐Charro, Bunge, & Guy, ). Ideally, skin testing addressing questions involving humans should be conducted on human skin.…”

Section: Introductionmentioning

confidence: 99%

Histological and functional comparisons of four anatomical regions of porcine skin with human abdominal skin

Richardson

Loewa

et al. 2019

Anat Histol Embryol

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Because of the shortage of human skin for research purposes, porcine skin has been used as a model of human skin. The aim of this study was to identify the region of German Landrace pig skin that could be used as the best possible substitute for human abdominal skin. Porcine samples were collected from the ear, flank, back and caudal abdomen; human abdominal skin samples were excised during plastic surgery. Histological and ultrastructural assessments were carried out on the epidermis and dermis, with emphasis on the dermo‐epidermal interface length, dermo‐epidermal thickness ratio as well as densities of; hair follicles, arrector pili muscles, blood vessels and sweat glands. In the pig, the barrier function of the four anatomical regions was assessed. Results showed that both histologically and ultrastructurally, all four regions of porcine skin were similar to human skin. These include the shapes of keratinocytes, structure of cell contacts and presence of Weibel Palade bodies in endothelial cells. Other parameters such as the thickness of epidermis, the thickness of stratum basale, spinosum and granulosum and the number of cell layers in the stratum corneum were similar in human abdominal and in all four regions of porcine skin. However, there were also significant differences especially in the thickness of the stratum corneum, the dermo‐epidermal interface length and the blood vessel density.

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Section: Introductionmentioning

confidence: 99%

Histological and functional comparisons of four anatomical regions of porcine skin with human abdominal skin

Richardson

Loewa

et al. 2019

Anat Histol Embryol

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“…This is often due to their prolonged retention and potential systemic toxicity post IV administration. As a result, several researchers are looking toward utilizing topical approaches to avoid negative systemic delivery effects [ 24 , 31 , 47 , 52 , 53 , 54 , 55 , 56 , 57 , 58 ]. Our topical approach of administering nanoparticles directly to the resected tissue, investigated here, supports the idea that our tumor targeting nanoparticles can still provide both sensitive and specific information between tumor and normal adjacent tissue specimens post resection.…”

Section: Discussionmentioning

confidence: 99%

A Raman Imaging Approach Using CD47 Antibody-Labeled SERS Nanoparticles for Identifying Breast Cancer and Its Potential to Guide Surgical Resection

et al. 2018

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Raman spectroscopic imaging has shown great promise for improved cancer detection and localization with the use of tumor targeting surface enhanced Raman scattering (SERS) nanoparticles. With the ultrasensitive detection and multiplexing capabilities that SERS imaging has to offer, scientists have been investigating several clinical applications that could benefit from this unique imaging strategy. Recently, there has been a push to develop new image-guidance tools for surgical resection to help surgeons sensitively and specifically identify tumor margins in real time. We hypothesized that SERS nanoparticles (NPs) topically applied to breast cancer resection margins have the potential to provide real-time feedback on the presence of residual cancer in the resection margins during lumpectomy. Here, we explore the ability of SERS nanoparticles conjugated with a cluster of differentiation-47 (CD47) antibody to target breast cancer. CD47 is a cell surface receptor that has recently been shown to be overexpressed on several solid tumor types. The binding potential of our CD47-labeled SERS nanoparticles was assessed using fluorescence assisted cell sorting (FACS) on seven different human breast cancer cell lines, some of which were triple negative (negative expression of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER2)). Xenograft mouse models were also used to assess the ability of our Raman imaging system to identify tumor from normal tissue. A ratiometric imaging strategy was used to quantify specific vs. nonspecific probe binding, resulting in improved tumor-to-background ratios. FACS analysis showed that CD47-labeled SERS nanoparticles bound to seven different breast cancer cell lines at levels 12-fold to 70-fold higher than isotype control-labeled nanoparticles (p < 0.01), suggesting that our CD47-targeted nanoparticles actively bind to CD47 on breast cancer cells. In a mouse xenograft model of human breast cancer, topical application of CD47-targeted nanoparticles to excised normal and cancer tissue revealed increased binding of CD47-targeted nanoparticles on tumor relative to normal adjacent tissue. The findings of this study support further investigation and suggest that SERS nanoparticles topically applied to breast cancer could guide more complete surgical resection during lumpectomy.

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“…On the other hand, frozen then thawed specimens sprayed with SPiDER-βGal showed comparable or even higher fluorescence signal than fresh specimens. This may be due to freeze-thaw induced membrane damage with increased permeability and stabilization of the activated form SPiDER-βGal to intracellular proteins [ 10 ]. With either probe, although frozen tissue can be thawed and analyzed, temperature control is even more crucial than with fresh specimens.…”

Section: Discussionmentioning

confidence: 99%

“…Small molecule enzyme-activatable fluorescence probes are rapidly cleaved and activated by a single enzymatic reaction using targeted endopeptidases. Often this process occurs in less than an hour making it practical for clinical translation [ 9 , 10 ]. Activatable probes have been investigated in preclinical animal models and some of them have recently proceeded to clinical trials [ 11 – 13 ].…”

Section: Introductionmentioning

confidence: 99%

“…β-Gal and GGT are overexpressed in a variety of cancers, and are potential candidates for intraoperative enzyme activated probes. In a previous report, we described the fluorescence signal and the kinetics generated by HMRef-βGal and SPiDER-βGal both activated by β-galactosidase and gGlu-HMRG which is activated by GGT in ovarian cancer cell lines at 37°C [ 10 ]. HMRef-βGal, the original probe [ 19 ], behaved with similar kinetics to gGlu-HMRG probe at this temperature although the fluorescence signal of HMRef-βGal was much lower than that of gGlu-HMRG.…”

Section: Introductionmentioning

confidence: 99%

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Pitfalls on sample preparation for ex vivo imaging of resected cancer tissue using enzyme-activatable fluorescent probes

et al. 2018

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In vivo and ex vivo fluorescence imaging-assisted surgery can aid in determining the margins of tumors during surgical resection. While a variety of fluorescent probes have been proposed for this task, small molecule enzyme-activatable fluorescent probes are ideal for this application. They are quickly activated at tumor sites and result in bright signal with little background, resulting in high sensitivity. Testing in resected specimens, however, can be difficult. Enzymes are usually stable after freezing and thawing but catalytic reactions are generally temperature-dependent. Therefore, tissue sample temperature should be carefully considered. In this study two enzyme activatable probes, γ-glutamylhydroxymethyl rhodamine green (gGlu-HMRG) that reacted with γ-glutamyltransferase and SPiDER-βGal that reacted with β-galactosidase, were employed to determine the effects of temperature on fluorescence signal kinetics in both fresh and frozen and then thawed ex vivo experimental ovarian cancer tissue samples. The results suggest γ-glutamyltransferase was less sensitive to temperature than β-galactosidase. Fresh samples showed higher fluorescence signals of gGlu-HMRG compared with thawed samples likely because the freeze-thaw cycle decreased the rate of internalization of the activated probe into the lysosome. In contrast, no significant difference of SPiDER-βGal fluorescence signal was observed between fresh and frozen tissues. In conclusion, although imaging of fresh samples at 37°C is the best condition for both probes, successful imaging with gGlu-HMRG could be achieved even at room temperature with thawed samples. We demonstrate that temperature regulation and tissue handling of resected tissue are two pitfalls that may influence ex vivo imaging signals with enzyme-activatable fluorescent probes.

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A topically-sprayable, activatable fluorescent and retaining probe, SPiDER-βGal for detecting cancer: Advantages of anchoring to cellular proteins after activation

Cited by 20 publications

References 32 publications

Histological and functional comparisons of four anatomical regions of porcine skin with human abdominal skin

Histological and functional comparisons of four anatomical regions of porcine skin with human abdominal skin

A Raman Imaging Approach Using CD47 Antibody-Labeled SERS Nanoparticles for Identifying Breast Cancer and Its Potential to Guide Surgical Resection

Pitfalls on sample preparation for ex vivo imaging of resected cancer tissue using enzyme-activatable fluorescent probes

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