Multi-Modality Therapeutics with Potent Anti-Tumor Effects: Photochemical Internalization Enhances Delivery of the Fusion Toxin scFvMEL/rGel
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{"title"=>"Multi-modality therapeutics with potent anti-tumor effects: Photochemical internalization enhances delivery of the fusion toxin scFvMEL/rGel", "type"=>"journal", "authors"=>[{"first_name"=>"Pål K.", "last_name"=>"Selbo", "scopus_author_id"=>"6603234454"}, {"first_name"=>"Michael G.", "last_name"=>"Rosenblum", "scopus_author_id"=>"7202183770"}, {"first_name"=>"Laurence H.", "last_name"=>"Cheung", "scopus_author_id"=>"7102302741"}, {"first_name"=>"Wendy", "last_name"=>"Zhang", "scopus_author_id"=>"35096887700"}, {"first_name"=>"Kristian", "last_name"=>"Berg", "scopus_author_id"=>"7401938257"}], "year"=>2009, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-68949213748", "pmid"=>"19690617", "sgr"=>"68949213748", "doi"=>"10.1371/journal.pone.0006691", "isbn"=>"1932-6203 (Electronic)\\n1932-6203 (Linking)", "issn"=>"19326203", "pui"=>"355130368"}, "id"=>"dbb82e38-a499-3fd6-9b70-016fcb6c8634", "abstract"=>"BACKGROUND: There is a need for drug delivery systems (DDS) that can enhance cytosolic delivery of anti-cancer drugs trapped in the endo-lysosomal compartments. Exposure of cells to specific photosensitizers followed by light exposure (photochemical internalization, PCI) results in transfer of agents from the endocytic compartment into the cytosol.\\n\\nMETHODOLOGY AND PRINCIPAL FINDINGS: The recombinant single-chain fusion construct scFvMEL/rGel is composed of an antibody targeting the progenitor marker HMW-MAA/NG2/MGP/gp240 and the highly effective toxin gelonin (rGel). Here we demonstrate enhanced tumor cell selectivity, cytosolic delivery and anti-tumor activity by applying PCI of scFvMEL/rGel. PCI performed by light activation of cells co-incubated with scFvMEL/rGel and the endo-lysosomal targeting photosensitizers AlPcS(2a) or TPPS(2a) resulted in enhanced cytotoxic effects against antigen-positive cell lines, while no differences in cytotoxicity between the scFvMEL/rGel and rGel were observed in antigen-negative cells. Mice bearing well-developed melanoma (A-375) xenografts (50-100 mm(3)) were treated with PCI of scFvMEL/rGel. By 30 days after injection, approximately 100% of mice in the control groups had tumors>800 mm(3). In contrast, by day 40, 50% of mice in the PCI of scFvMEL/rGel combination group had tumors<800 mm(3) with no increase in tumor size up to 110 days. PCI of scFvMEL/rGel resulted in a synergistic effect (p<0.05) and complete regression (CR) in 33% of tumor-bearing mice (n = 12).\\n\\nCONCLUSIONS/SIGNIFICANCE: This is a unique demonstration that a non-invasive multi-modality approach combining a recombinant, targeted therapeutic such as scFvMEL/rGel and PCI act in concert to provide potent in vivo efficacy without sacrificing selectivity or enhancing toxicity. The present DDS warrants further evaluation of its clinical potential.", "link"=>"http://www.mendeley.com/research/multimodality-therapeutics-potent-antitumor-effects-photochemical-internalization-enhances-delivery", "reader_count"=>28, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Researcher"=>6, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>6, "Student > Master"=>8, "Other"=>3, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Researcher"=>6, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>6, "Student > Master"=>8, "Other"=>3, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>8, "Medicine and Dentistry"=>11, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Physics and Astronomy"=>1, "Psychology"=>1, "Chemistry"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>11}, "Chemistry"=>{"Chemistry"=>1}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Psychology"=>{"Psychology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>8}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"United States"=>2, "Poland"=>1, "Germany"=>1}, "group_count"=>3}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/887724"], "description"=>"<p>The values are based on paired log rank analyses of the time for the A-375 tumors to reach 800 mm<sup>3</sup>.</p>", "links"=>[], "tags"=>["analyses", "differences", "therapeutic", "regimens", "a-375"], "article_id"=>558178, "categories"=>["Cancer", "Biochemistry", "Biophysics"], "users"=>["Pål K. Selbo", "Michael G. Rosenblum", "Lawrence H. Cheung", "Wendy Zhang", "Kristian Berg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006691.t002", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Statistical_significance_analyses_p_values_to_reveal_differences_in_therapeutic_effect_between_various_treatment_regimens_on_the_A_375_tumors_/558178", "title"=>"Statistical significance analyses (p-values) to reveal differences in therapeutic effect between various treatment regimens on the A-375 tumors.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2009-08-19 02:16:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/887415"], "description"=>"<p>The cells were incubated with 5.0 µg/ml AlPcS<sub>2a</sub>+/−16.5 nM scFvMEL/rGel or 16.5 nM rGel for 18 hours and then washed twice and chased in drug-free medium 4 h prior to red light exposure. MTT assay was performed 48 hours post light. Experiments with triplicates were reproduced at least twice. Bars, SE.</p>", "links"=>[], "tags"=>["ma11", "carcinoma", "cells", "u87", "malignant", "glioma"], "article_id"=>557860, "categories"=>["Cancer", "Biochemistry", "Biophysics"], "users"=>["Pål K. Selbo", "Michael G. Rosenblum", "Lawrence H. Cheung", "Wendy Zhang", "Kristian Berg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006691.g003", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_PCI_of_scFvMEL_rGel_in_MA11_breast_carcinoma_cells_A_and_U87_malignant_glioma_cells_B_/557860", "title"=>"PCI of scFvMEL/rGel in MA11 breast carcinoma cells (A) and U87 malignant glioma cells (B).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2009-08-19 02:11:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/887474"], "description"=>"<p>A, Cells were treated with either rGel or scFvMEL/rGel for 18 h with increasing toxin concentrations. B, AlPcS<sub>2a</sub>-PCI of scFvMEL/rGel or rGel in T24 cells. The cells were treated and assessed as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0006691#pone-0006691-g003\" target=\"_blank\">figure 3</a>. Bars, SE.</p>", "links"=>[], "tags"=>["experiments", "gp240-negative", "bladder", "cancer"], "article_id"=>557926, "categories"=>["Cancer", "Biochemistry", "Biophysics"], "users"=>["Pål K. Selbo", "Michael G. Rosenblum", "Lawrence H. Cheung", "Wendy Zhang", "Kristian Berg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006691.g004", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Control_experiments_in_gp240_negative_bladder_cancer_cell_line_T24_/557926", "title"=>"Control experiments in gp240-negative bladder cancer cell line T24.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2009-08-19 02:12:06"}
  • {"files"=>["https://ndownloader.figshare.com/files/887123"], "description"=>"<p>A375-GFP cells were incubated for 18 h with 5 µg/ml AlPcS<sub>2a</sub>, washed twice and chased in drug-free medium for 4 h. 30 min prior to microscopy, cells were incubated with 5 µg/ml Hoechst 33342. A, Fluorescence of EGFP, Hoechst 33342 and AlPcS<sub>2a</sub> and merged (E+H+A) in vital cells prior to (upper panels) and 1 min post 10 sec of microscopy red light exposure (590–650 nm) with respectively merged micrographs and corresponding phase contrast (mid panels). Structure formula of AlPcS<sub>2a</sub> in left lower panel. B, A-375 cells were co-incubated for 18 h with 80 nM scFvMEL/rGel and 5 µg/ml AlPcS<sub>2a</sub>, washed twice and chased in drug-free medium for 4 h prior to light exposure and further incubated at 37°C for 30 min prior to ICC detection of rGel. Right panel, negative controls included rabbit IgG isotype.</p>", "links"=>[], "tags"=>["endo-lysosomal", "cytosolic", "photosensitizer"], "article_id"=>557577, "categories"=>["Cancer", "Biochemistry", "Biophysics"], "users"=>["Pål K. Selbo", "Michael G. Rosenblum", "Lawrence H. Cheung", "Wendy Zhang", "Kristian Berg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006691.g001", "stats"=>{"downloads"=>1, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Photochemical_mediated_endo_lysosomal_release_and_cytosolic_delivery_of_photosensitizer_and_scFvMEL_rGel_/557577", "title"=>"Photochemical-mediated endo-lysosomal release and cytosolic delivery of photosensitizer and scFvMEL/rGel.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2009-08-19 02:06:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/887534"], "description"=>"<p>A, Kaplan Meier plot of treatment groups as indicated in the figure. Nude mice with s.c. growing A-375 (50–100 mm3) tumors were given AlPcS<sub>2a</sub> (5 mg/kg i.p.) with or without scFvMEL/rGel (2 mg/kg i.v) administered 48 and 24 hours prior to laser light exposure (day 0), respectively. The light dose was 20 J/cm<sup>2</sup> with an irradiance of 100 mW/cm<sup>2</sup>. n, number of animals in each group. B, Weights of the mice treated with PCI of scFvMEL/rGel as compared with non-treated animals. Range of weights at treatment start were 22–29 g. Weight data for the other groups are shown in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0006691#pone.0006691.s006\" target=\"_blank\">Figure S6</a>.</p>", "links"=>[], "tags"=>["a-375"], "article_id"=>557996, "categories"=>["Cancer", "Biochemistry", "Biophysics"], "users"=>["Pål K. Selbo", "Michael G. Rosenblum", "Lawrence H. Cheung", "Wendy Zhang", "Kristian Berg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006691.g005", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_PCI_of_scFvMEL_rGel_in_A_375_xenografts_/557996", "title"=>"PCI of scFvMEL/rGel in A-375 xenografts.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2009-08-19 02:13:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/887767"], "description"=>"<p>Mean time for A-375 tumors to reach the endpoint of 800 mm<sup>3</sup> after various specified treatments based on Kaplan-Meier survival analysis in SPSS where all animals in the groups are included. SE represents standard error.</p>", "links"=>[], "tags"=>["a-375", "tumors", "endpoint", "800"], "article_id"=>558224, "categories"=>["Cancer", "Biochemistry", "Biophysics"], "users"=>["Pål K. Selbo", "Michael G. Rosenblum", "Lawrence H. Cheung", "Wendy Zhang", "Kristian Berg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006691.t001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mean_time_for_A_375_tumors_to_reach_the_endpoint_of_800_mm_3_/558224", "title"=>"Mean time for A-375 tumors to reach the endpoint of 800 mm<sup>3</sup>.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2009-08-19 02:17:04"}
  • {"files"=>["https://ndownloader.figshare.com/files/439797", "https://ndownloader.figshare.com/files/439832", "https://ndownloader.figshare.com/files/439867", "https://ndownloader.figshare.com/files/439949", "https://ndownloader.figshare.com/files/439995", "https://ndownloader.figshare.com/files/440063"], "description"=>"<div><h3>Background</h3><p>There is a need for drug delivery systems (DDS) that can enhance cytosolic delivery of anti-cancer drugs trapped in the endo-lysosomal compartments. Exposure of cells to specific photosensitizers followed by light exposure (photochemical internalization, PCI) results in transfer of agents from the endocytic compartment into the cytosol.</p><h3>Methodology and Principal Findings</h3><p>The recombinant single-chain fusion construct scFvMEL/rGel is composed of an antibody targeting the progenitor marker HMW-MAA/NG2/MGP/gp240 and the highly effective toxin gelonin (rGel). Here we demonstrate enhanced tumor cell selectivity, cytosolic delivery and anti-tumor activity by applying PCI of scFvMEL/rGel. PCI performed by light activation of cells co-incubated with scFvMEL/rGel and the endo-lysosomal targeting photosensitizers AlPcS<sub>2a</sub> or TPPS<sub>2a</sub> resulted in enhanced cytotoxic effects against antigen-positive cell lines, while no differences in cytotoxicity between the scFvMEL/rGel and rGel were observed in antigen-negative cells. Mice bearing well-developed melanoma (A-375) xenografts (50–100 mm<sup>3</sup>) were treated with PCI of scFvMEL/rGel. By 30 days after injection, ∼100% of mice in the control groups had tumors>800 mm<sup>3</sup>. In contrast, by day 40, 50% of mice in the PCI of scFvMEL/rGel combination group had tumors<800 mm<sup>3</sup> with no increase in tumor size up to 110 days. PCI of scFvMEL/rGel resulted in a synergistic effect (p<0.05) and complete regression (CR) in 33% of tumor-bearing mice (n = 12).</p><h3>Conclusions/Significance</h3><p>This is a unique demonstration that a non-invasive multi-modality approach combining a recombinant, targeted therapeutic such as scFvMEL/rGel and PCI act in concert to provide potent <em>in vivo</em> efficacy without sacrificing selectivity or enhancing toxicity. The present DDS warrants further evaluation of its clinical potential.</p></div>", "links"=>[], "tags"=>["multi-modality", "therapeutics", "potent", "anti-tumor", "photochemical", "internalization", "enhances", "fusion", "toxin"], "article_id"=>146700, "categories"=>["Cancer", "Biochemistry", "Biophysics"], "users"=>["Pål K. Selbo", "Michael G. Rosenblum", "Lawrence H. Cheung", "Wendy Zhang", "Kristian Berg"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0006691.s001", "https://dx.doi.org/10.1371/journal.pone.0006691.s002", "https://dx.doi.org/10.1371/journal.pone.0006691.s003", "https://dx.doi.org/10.1371/journal.pone.0006691.s004", "https://dx.doi.org/10.1371/journal.pone.0006691.s005", "https://dx.doi.org/10.1371/journal.pone.0006691.s006"], "stats"=>{"downloads"=>4, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Multi_Modality_Therapeutics_with_Potent_Anti_Tumor_Effects_Photochemical_Internalization_Enhances_Delivery_of_the_Fusion_Toxin_scFvMEL_rGel/146700", "title"=>"Multi-Modality Therapeutics with Potent Anti-Tumor Effects: Photochemical Internalization Enhances Delivery of the Fusion Toxin scFvMEL/rGel", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2009-08-19 01:51:40"}
  • {"files"=>["https://ndownloader.figshare.com/files/887287"], "description"=>"<p>A, Viability of A375-GFP cells subjected to PCI of scFvMEL/rGel (red triangles), PDT (black dots) or scFvMEL/rGel only (green squares). Cells were incubated with 5 µg/ml AlPcS<sub>2a</sub> with or without 100 nM scFvMEL/rGel for 18 h, followed by 3 times wash and 4 h chase in drug-free medium prior to exposure to increasing light exposure of cells (15 minutes = 1.35 J/cm<sup>2</sup>). Cytotoxicity was assessed by the MTT assay 48 h post light exposure. Bars, SD. B, Fluorescence of EGFP and Hoechst 33342 in an untreated A375-GFP cell. C, Phase contrast of a cell treated with PCI of scFvMEL/rGel (LD90, 0.54 J/cm<sup>2</sup>) with corresponding fluorescence of AlPcS<sub>2a</sub> (D), Hoechst 33342 (E), EGFP (F) and a merge of D,E and F (G) 48 h post light exposure. H, A375-GFP cells 24 and 48 hours post either PDT (LD50, 1.35 J/cm<sup>2</sup>) or PCI of scFvMEL/rGel (LD90, 0.54 J/cm<sup>2</sup>) as compared to untreated control cells (Ctr).</p>", "links"=>[], "tags"=>["cytotoxicity", "egfp", "signals", "pci", "a375-gfp", "melanoma"], "article_id"=>557737, "categories"=>["Cancer", "Biochemistry", "Biophysics"], "users"=>["Pål K. Selbo", "Michael G. Rosenblum", "Lawrence H. Cheung", "Wendy Zhang", "Kristian Berg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006691.g002", "stats"=>{"downloads"=>1, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Enhancement_of_cytotoxicity_and_reduction_of_EGFP_signals_by_PCI_of_scFvMEL_rGel_in_A375_GFP_melanoma_cells_/557737", "title"=>"Enhancement of cytotoxicity and reduction of EGFP signals by PCI of scFvMEL/rGel in A375-GFP melanoma cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2009-08-19 02:08:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/887651"], "description"=>"<p>Two mice (#1 and #2) with ∼75 mm3 A-375 tumors at day 0 were further assessed at day 1, 5 and 19 post PCI. Both mice obtained CR and excellent wound repair after PCI of scFvMEL/rGel (still at day 110) as compared to a untreated control mouse (Ctr) having a tumor which reached >1000 mm3 at day 19 (∼13-fold tumor increase compared to day 0).</p>", "links"=>[], "tags"=>["regeneration", "pci"], "article_id"=>558109, "categories"=>["Cancer", "Biochemistry", "Biophysics"], "users"=>["Pål K. Selbo", "Michael G. Rosenblum", "Lawrence H. Cheung", "Wendy Zhang", "Kristian Berg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006691.g006", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Tumor_response_and_skin_regeneration_post_PCI_of_scFvMEL_rGel_/558109", "title"=>"Tumor response and skin regeneration post PCI of scFvMEL/rGel.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2009-08-19 02:15:09"}

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Relative Metric

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