183 results on '"Horak, Ivan D."'
Search Results
152. Gene-specific repair in human CD4+ lymphocytes reflects transcription and proliferation
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Bartośova, Zdena, primary, Pirśel, Miroslav, additional, Reinhold, William, additional, Stetler-Stevenson, Maryalice, additional, Zajac-Kaye, Maria, additional, May, Alfred, additional, Horak, Ivan D, additional, and Bohr, Vilhelm A, additional
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- 1996
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153. Non-amine based analogs of lavendustin A as protein-tyrosine kinase inhibitors
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Smyth, Mark S., primary, Stefanova, Irena, additional, Hartmann, Frank, additional, Horak, Ivan D., additional, Osherov, Nir, additional, Levitzki, Alexander, additional, and Burke, Terrence R., additional
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- 1993
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154. Hydroxylated 2-(5'-salicyl)naphthalenes as protein-tyrosine kinase inhibitors
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Smyth, Mark S., primary, Stefanova, Irena, additional, Horak, Ivan D., additional, and Burke, Terrence R., additional
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- 1993
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155. Bicyclic compounds as ring-constrained inhibitors of protein-tyrosine kinase p56lck
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Burke, Terrence R., primary, Lim, Benjamin, additional, Marquez, Victor E., additional, Li, Zhen Hong, additional, Bolen, Joseph B., additional, Stefanova, Irena, additional, and Horak, Ivan D., additional
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- 1993
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156. Arylamides of hydroxylated isoquinolines as protein-tyrosine kinase inhibitors
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Burke, Terrence R., primary, Ford, Harry, additional, Osherov, Nir, additional, Levitzki, Alexander, additional, Stefanova, Irena, additional, Horak, Ivan D., additional, and Marquez, Victor E., additional
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- 1992
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157. Lymphoblastic lymphoma presenting in cutaneous sites
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Sander, Christian A., primary, Medeiros, L. Jeffrey, additional, Abruzzo, Lynne V., additional, Horak, Ivan D., additional, and Jaffe, Elaine S., additional
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- 1991
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158. Clinical-Scale Radiolabeling of a Humanized Anticarcinoembryonic Antigen Monoclonal Antibody, hMN-14, with Residualizing 131I for Use in Radioimmunotherapy.
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Govindan, Serengulam V., Griffiths, Gary L., Stein, Rhona, Andrews, Philip, Sharkey, Robert M., Hansen, Hans J., Horak, Ivan D., and Goldenberg, David M.
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- 2005
159. Reagents and Methods for PET Using Bispecific Antibody Pretargeting and 68Ga-Radiolabeled Bivalent Hapten-Peptide-Chelate Conjugates.
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Griffiths, Gary L., Chien-Hsing Chang, McBride, William J., Rossi, Edmund A., Sheerin, Agatha, Tejada, German R., Karacay, Habibe, Sharkey, Robert M., Horak, Ivan D., Hansen, Hans J., and Goldenberg, David M.
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- 2004
160. Effective therapy of human lymphoma xenografts with a novel recombinant ribonuclease/anti-CD74 humanized IgG4antibody immunotoxin
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Chang, Chien-Hsing, Sapra, Puja, Vanama, Sailaja S., Hansen, Hans J., Horak, Ivan D., and Goldenberg, David M.
- Abstract
Ranpirnase (Rap) is a cytotoxic ribonuclease (RNase) isolated from frog oocytes. Here we describe high antitumor activity of a novel immunotoxin, 2L-Rap-hLL1-γ4P, composed of 2 Rap molecules, each fused to the N terminus of the light chain of hLL1, an internalizing anti-CD74 humanized antibody. To reduce unwanted side effects, the constant region of hLL1 was changed from γ1 to γ4 and further to γ4P by replacing serine228to proline to prevent the formation of a half immunoglobulin G (IgG) common for IgG4. In vitro, 2L-Rap-hLL1-γ4P retained RNase activity, specific binding to CD74, and was significantly more potent against CD74+cell lines (Daudi, Raji, and MC/CAR) than naked hLL1. In vivo, the pharmacokinetic profile of 2L-Rap-hLL1-γ4P was similar to that of naked hLL1. The maximum tolerated dose of 2L-Rap-hLL1-γ4P in severe combined immunodeficient mice (SCID) or BALB/c mice was 50 μg per mouse. In Raji and Daudi Burkitt lymphoma xenograft models, treatment with a single 5 to 50 μg dose of 2L-Rap-hLL1-γ4P, given as early or delayed treatment, resulted in cures of most animals. Treatment with 2L-Rap-hLL1-γ4P was significantly better than all controls, including saline, naked hLL1, and nonspecific immunotoxin. In conclusion, 2L-Rap-hLL1-γ4P demonstrated excellent in vitro and in vivo efficacy and thus merits further consideration as a therapeutic for CD74+tumors.
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- 2005
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161. Gene-specific repair in human CD4+lymphocytes reflects transcription and proliferation
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Bartośova, Zdena, Pirśel, Miroslav, Reinhold, William, Stetler-Stevenson, Maryalice, Zajac-Kaye, Maria, May, Alfred, Horak, Ivan D, and Bohr, Vilhelm A
- Abstract
We have measured the gene-specific repair of ultraviolet irradiation (UV)-induced cyclobutane pyrimidine dimers (CPD) in freshly isolated human peripheral blood CD4+T-lymphocytes. Two populations of CD4+lymphocytes were assayed: resting and proliferating cells. DNA repair was assessed in the essential gene dihydrofolate reductase (DHFR) as well as in each of its strands, in the proliferation inducible c-mycgene and in the inactive δ-globin gene. Transcription rates in these genes were determined by nuclear run-on assay in the two cell populations. The rate of DHFR transcription increased 10-fold from resting to proliferating lymphocytes. Transcripts from c-mycwere present only in proliferating cells, and we detected no δ-globin transcripts in either cell population. During the 24-h period after UV irradiation, there was little or no repair in any of the genes in the resting cells; there was some repair in the transcribed strand of the DHFR gene, but no repair in its nontranscribed strand. In the proliferating cells where the transcription of DHFR was much increased, the repair was efficient. The δ-globin gene was not expressed in either cell population, but it was more efficiently repaired in the proliferating than in the resting cells. We suggest that the gene-specific repair activity in CD4+lymphocytes can reflect the proliferative state of the cells as well as the transcriptional state of the gene.
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- 1996
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162. Efficacy of Sym004 in Patients With Metastatic Colorectal Cancer With Acquired Resistance to Anti-EGFR Therapy and Molecularly Selected by Circulating Tumor DNA Analyses
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Montagut, Clara, Argilés, Guillem, Ciardiello, Fortunato, Poulsen, Thomas T., Dienstmann, Rodrigo, Kragh, Michael, Kopetz, Scott, Lindsted, Trine, Ding, Cliff, Vidal, Joana, Clausell-Tormos, Jenifer, Siravegna, Giulia, Sánchez-Martín, Francisco J., Koefoed, Klaus, Pedersen, Mikkel W., Grandal, Michael M., Dvorkin, Mikhail, Wyrwicz, Lucjan, Rovira, Ana, Cubillo, Antonio, Salazar, Ramon, Desseigne, Françoise, Nadal, Cristina, Albanell, Joan, Zagonel, Vittorina, Siena, Salvatore, Fumi, Guglielmo, Rospo, Giuseppe, Nadler, Paul, Horak, Ivan D., Bardelli, Alberto, and Tabernero, Josep
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163. In Vitroand In VivoTargeting and Therapy of an Antibody-Drug Conjugate (IMMU-110) in B-Cell Malignancies.
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Sapra, Puja, Stein, Rhona, Pickett, Jennifer, Govindan, Serengulam V., Cardillo, Thomas M., Damoci, Christopher, Sheerin, Agatha, Hansen, Hans J., Horak, Ivan D., Griffiths, Gary L., and Goldenberg, David M.
- Abstract
IMMU-110 is a drug immunoconjugate comprised of doxorubicin (DOX) conjugated to the humanized anti-CD74 monoclonal antibody (mAb), hLL1, at a DOX:mAb (mol/mol) ratio of 8:1. CD74 is a rapidly internalizing type-II transmembrane chaperone molecule associated with HLA-DR, and has high expression on human non-Hodgkin's lymphoma (NHL) and multiple myeloma (MM) clinical specimens and cell lines. Here, we investigated the in vitroand in vivoefficacy of IMMU-110 in xenograft models of human NHL (Raji, Daudi) and MM (MC/CAR). In vitrocell binding of IMMU-110 with the CD74-positive cells was significantly higher than that of a non-specific isotype-matched mAb-DOX conjugate (DOX conjugated to a mAb against epithelial glycoprotein-1; DOX-hRS7), and was similar to that of naked hLL1. Both IMMU-110 and naked hLL1 bound CD74 with subnanomolar affinity. The in vitrocytotoxicity of IMMU-110 was significantly higher than non-specific antibody-DOX conjugate, DOX-hRS7, and was similar to free DOX in MC/CAR, Raji or Daudi human Burkitt's lymphoma cells. In CD74-negative cell lines, IMMU-110 was significantly less toxic than free DOX, having similar cytotoxicity to DOX-hRS7. In vivo, IMMU-110 displayed a pharmacokinetic and biodistribution profile almost identical to that of hLL1 mAb. Both hLL1 mAb and IMMU-110 had a biphasic clearance from the circulation; the α and β half-life (t1/2) of IMMU-110 were 4.6 h and 157.9 h, respectively, and those of hLL1 were 5.4 h and 151.5 h, respectively. In biodistribution studies, no significant difference was observed between IMMU-110 and naked hLL1 with regards to normal tissue uptake. Neither IMMU-110 nor naked hLL1 mAb had a significant association with any normal body tissue. In therapy experiments, a single i.v. protein dose of 350 μg IMMU-110, injected 5 days after implantation of MC/CAR cells in SCID mice, resulted in curing 70% of the animals. Similar cure rates were observed when treatment with IMMU-110 was given 10 days after transplantation of MC/CAR cells. In the Raji xenograft model, 100% of animals were cured with a single protein dose of 120 μg IMMU-110, injected 5 days after implantation of cells. In survival studies, the efficacy of IMMU-110 was significantly better than naked hLL1, the combination of naked hLL1 and free DOX, or of a non-specific antibody-DOX conjugate, DOX-hRS7. In a tolerability study in SCID mice, no toxic effect of IMMU-110 was observed even at the highest dose tested (2.5 mg /mouse). In conclusion, treatment of B-cell lymphoma and myeloma xenograft models with single injections of IMMU-110 resulted in high levels of response and long-term survivors. IMMU-110 is being further developed as a potential therapeutic for the treatment of CD74-positive tumors.
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- 2004
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164. Sym004, a Novel EGFR Antibody Mixture, Can Overcome Acquired Resistance to Cetuximab.
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Iida, Mari, Brand, Toni M., Starr, Megan M., Chunrong Li, Huppert, Evan J., Luthar, Neha, Pedersen, Mikkel W., Horak, Ivan D., Kragh, Michael, and Wheeler, Deric L.
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EPIDERMAL growth factor receptors , *MONOCLONAL antibodies , *CETUXIMAB , *LUNG cancer , *CANCER cells - Abstract
The epidermal growth factor receptor (EGFR) is a central regulator of tumor progression in a variety of human cancers. Cetuximab is an anti-EGFR monoclonal antibody that has been approved for head and neck and colorectal cancer treatment, but many patients treated with cetuximab don't respond or eventually acquire resistance. To determine how tumor cells acquire resistance to cetuximab, we previously developed a model of acquired resistance using the non-small cell lung cancer line NCI-H226. These cetuximab-resistant (CtxR) cells exhibit increased steady-state EGFR expression secondary to alterations in EGFR trafficking and degradation and, further, retained dependence on EGFR signaling for enhanced growth potential. Here, we examined Sym004, a novel mixture of antibodies directed against distinct epitopes on the extracellular domain of EGFR, as an alternative therapy for CtxR tumor cells. Sym004 treatment of CtxR clones resulted in rapid EGFR degradation, followed by robust inhibition of cell proliferation and down-regulation of several mitogen-activated protein kinase pathways. To determine whether Sym004 could have therapeutic benefit in vivo, we established de novo CtxR NCI-H226 mouse xenografts and subsequently treated CtxR tumors with Sym004. Sym004 treatment of mice harboring CtxR tumors resulted in growth delay compared to mice continued on cetuximab. Levels of total and phospho-EGFR were robustly decreased in Ctx tumors treated with Sym004. Immunohistochemical analysis of these Sym004-treated xenograft tumors further demonstrated decreased expression of Ki67, and phospho-rpS6, as well as a modest increase in cleaved caspase-3. These results indicate that Sym004 may be an effective targeted therapy for CtxR tumors. [ABSTRACT FROM AUTHOR]
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- 2013
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165. Carcinoembryonic antigen antibody inhibits lung metastasis and augments chemotherapy in a human colonic carcinoma xenograft.
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Blumenthal, Rosalyn D., Osorio, Lou, Hayes, Marianne K., Horak, Ivan D., Hansen, Hans J., and Goldenberg, David M.
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TUMORS , *CELL growth , *CANCER cells , *CELL culture , *CELL death , *CELL proliferation - Abstract
Purpose: In addition to its use as a blood marker for many carcinomas, elevated expression of carcinoembryonic antigen (CEA, CD66e, CEACAM5) has been implicated in various biological aspects of neoplasia, especially tumor cell adhesion, metastasis, the blocking of cellular immune mechanisms, and having antiapoptosis functions. However, it is not known if treatment with anti-CEA antibodies can affect tumor metastasis or alter the effects of cytotoxic drugs. Methods: In vitro, human colon cancer cell lines were treated with anti-CEA MAb IgG1, hMN-14 (labetuzumab), to assess direct effects on proliferation, as well as antibody-dependent cellular cytotoxicity (ADCC), and complement-dependent cytotoxicity (CDC). In vivo studies were undertaken in nude mice bearing s.c. (local growth) or i.v. (metastatic model) GW-39 and LS174T human colon cancer grafts, to evaluate the MAb alone and in combination with either CPT-11 or 5-fluorouracil (5FU). Results: In vitro, labetuzumab did not induce apoptosis, nor did it affect tumor cell proliferation directly or by CDC, but it did inhibit tumor cell proliferation by ADCC. In vivo, labetuzumab did not increase median survival in the GW-39 metastatic model unless the mice were pretreated with GM-CSF to increase their peripheral WBC counts; GM-CSF alone was ineffective. Also, if GW-39 tumors were pretreated with IFN-? to up-regulate CEA expression threefold prior to i.v. injection, labetuzumab significantly increased median survival of the mice. When nude mice received labetuzumab with CPT-11 or 5FU, median survival increased significantly as compared to the drug or antibody alone. Conclusions: Labetuzumab, a CEA-specific MAb, induces effector-cell function in vitro against CEA-positive colonic tumor cells, and also inhibits growth of lung metastasis when CEA expression is up-regulated or if peripheral WBCs are increased. The MAb also shows chemosensitizing properties. [ABSTRACT FROM AUTHOR]
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- 2005
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166. Efficacy of Sym004 in Patients With Metastatic Colorectal Cancer With Acquired Resistance to Anti-EGFR Therapy and Molecularly Selected by Circulating Tumor DNA Analyses
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Françoise Desseigne, Klaus Koefoed, Mikkel W. Pedersen, Rodrigo Dienstmann, C. Ding, Thomas Tuxen Poulsen, Trine Lindsted, Joana Vidal, Jenifer Clausell-Tormos, Michael M. Grandal, Salvatore Siena, Ana Rovira, Giulia Siravegna, Francisco J. Sánchez-Martín, Giuseppe Rospo, Guillem Argiles, Antonio Cubillo, Alberto Bardelli, Cristina Nadal, Michael Kragh, Clara Montagut, Ivan D. Horak, Lucjan Wyrwicz, Mikhail Dvorkin, Joan Albanell, Scott Kopetz, Guglielmo Fumi, Paul Nadler, Fortunato Ciardiello, Ramon Salazar, Josep Tabernero, Vittorina Zagonel, Montagut, Clara, Argilés, Guillem, Ciardiello, Fortunato, Poulsen, Thomas T, Dienstmann, Rodrigo, Kragh, Michael, Kopetz, Scott, Lindsted, Trine, Ding, Cliff, Vidal, Joana, Clausell-Tormos, Jenifer, Siravegna, Giulia, Sánchez-Martín, Francisco J, Koefoed, Klau, Pedersen, Mikkel W, Grandal, Michael M, Dvorkin, Mikhail, Wyrwicz, Lucjan, Rovira, Ana, Cubillo, Antonio, Salazar, Ramon, Desseigne, Françoise, Nadal, Cristina, Albanell, Joan, Zagonel, Vittorina, Siena, Salvatore, Fumi, Guglielmo, Rospo, Giuseppe, Nadler, Paul, Horak, Ivan D, Bardelli, Alberto, and Tabernero, Josep
- Subjects
Adult ,Male ,0301 basic medicine ,Oncology ,Cancer Research ,medicine.medical_specialty ,Colorectal cancer ,Population ,Loading dose ,Circulating Tumor DNA ,law.invention ,03 medical and health sciences ,0302 clinical medicine ,Randomized controlled trial ,law ,Internal medicine ,Biomarkers, Tumor ,medicine ,Humans ,Neoplasm Metastasis ,education ,Protein Kinase Inhibitors ,Survival analysis ,Aged ,Aged, 80 and over ,education.field_of_study ,business.industry ,Patient Selection ,Hazard ratio ,Antibodies, Monoclonal ,Correction ,Middle Aged ,medicine.disease ,Survival Analysis ,3. Good health ,ErbB Receptors ,Clinical trial ,Treatment Outcome ,030104 developmental biology ,Drug Resistance, Neoplasm ,030220 oncology & carcinogenesis ,Biomarker (medicine) ,Female ,Colorectal Neoplasms ,business - Abstract
Importance Acquired resistance to anti-EGFR therapy (epidermal growth factor receptor) is frequently due toRASandEGFRextracellular domain (ECD) mutations in metastatic colorectal cancer (mCRC). Some anti-EGFR–refractory patients retain tumor EGFR dependency potentially targetable by agents such as Sym004, which is a mixture of 2 nonoverlapping monoclonal antibodies targeting EGFR. Objective To determine if continuous blockade of EGFR by Sym004 has survival benefit. Design, Setting, and Participants Multicenter, phase 2, randomized, clinical trial comparing 2 regimens of Sym004 with investigator’s choice from March 6, 2014, through October 15, 2015. Circulating tumor DNA (ctDNA) was analyzed for biomarker and tracking clonal dynamics during treatment. Participants had wild-typeKRASexon 2 mCRC refractory to standard chemotherapy and acquired resistance to anti-EGFR monoclonal antibodies. Interventions Participants were randomly assigned in a 1:1:1 ratio to Sym004, 12 mg/kg/wk (arm A), Sym004, 9 mg/kg loading dose followed by 6 mg/kg/wk (arm B), or investigator’s choice of treatment (arm C). Main Outcomes and Measures Overall survival (OS). Secondary end points included preplanned exploratory biomarker analysis in ctDNA. Results A total of 254 patients were randomized (intent-to-treat [ITT] population) (median age, 63 [range, 34-91] years; 63% male; n = 160). Median OS in the ITT population was 7.9 months (95% CI, 6.5-9.9 months), 10.3 months (95% CI, 9.0-12.9 months), and 9.6 months (95% CI, 8.3-12.2 months) for arms A, B, and C, respectively (hazard ratio [HR], 1.31; 95% CI, 0.92-1.87 for A vs C; and HR, 0.97; 95% CI, 0.68-1.40 for B vs C). The ctDNA revealed high intrapatient genomic heterogeneity following anti-EGFR therapy. Sym004 effectively targetedEGFRECD-mutated cancer cells, and a decrease inEGFRECD ctDNA occurred in Sym004-treated patients. However, this did not translate into clinical benefit in patients withEGFRECD mutations, likely owing to co-occurring resistance mechanisms. A subgroup of patients was defined by ctDNA (RAS/BRAF/EGFRECD-mutation negative) associated with improved OS in Sym004-treated patients in arm B compared with arm C (median OS, 12.8 and 7.3 months, respectively). Conclusions and Relevance Sym004 did not improve OS in an unselected population of patients with mCRC and acquired anti-EGFR resistance. A prospective clinical validation of Sym004 efficacy in a ctDNA molecularly defined subgroup of patients with refractory mCRC is warranted. Trial Registration clinicaltrialsregister.eu Identifier:2013-003829-29
- Published
- 2018
167. An ERBB1-3 Neutralizing Antibody Mixture With High Activity Against Drug-Resistant HER2+ Breast Cancers With ERBB Ligand Overexpression.
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Schwarz, Luis J, Hutchinson, Katherine E, Rexer, Brent N, Estrada, Mónica Valeria, Gonzalez Ericsson, Paula I, Sanders, Melinda E, Dugger, Teresa C, Formisano, Luigi, Guerrero-Zotano, Angel, Red-Brewer, Monica, Young, Christian D, Lantto, Johan, Pedersen, Mikkel W, Kragh, Michael, Horak, Ivan D, and Arteaga, Carlos L
- Abstract
Background: Plasticity of the ERBB receptor network has been suggested to cause acquired resistance to anti-human epidermal growth factor receptor 2 (HER2) therapies. Thus, we studied whether a novel approach using an ERBB1-3-neutralizing antibody mixture can block these compensatory mechanisms of resistance.Methods: HER2+ cell lines and xenografts (n ≥ 6 mice per group) were treated with the ERBB1-3 antibody mixture Pan-HER, trastuzumab/lapatinib (TL), trastuzumab/pertuzumab (TP), or T-DM1. Downregulation of ERBB receptors was assessed by immunoblot analysis and immunohistochemistry. Paired pre- and post-T-DM1 tumor biopsies from patients (n = 11) with HER2-amplified breast cancer were evaluated for HER2 and P-HER3 expression by immunohistochemistry and/or fluorescence in situ hybridization. ERBB ligands were measured by quantitative reverse transcription polymerase chain reaction. Drug-resistant cells were generated by chronic treatment with T-DM1. All statistical tests were two-sided.Results: Treatment with Pan-HER inhibited growth and promoted degradation of ERBB1-3 receptors in a panel of HER2+ breast cancer cells. Compared with TL, TP, and T-DM1, Pan-HER induced a similar antitumor effect against established BT474 and HCC1954 tumors, but was superior to TL against MDA-361 xenografts (TL mean = 2026 mm 3 , SD = 924 mm 3 , vs Pan-HER mean = 565 mm 3 , SD = 499 mm 3 , P = .04). Pan-HER-treated BT474 xenografts did not recur after treatment discontinuation, whereas tumors treated with TL, TP, and T-DM1 did. Post-TP and post-T-DM1 recurrent tumors expressed higher levels of neuregulin-1 (NRG1), HER3 and P-HER3 (all P < .05). Higher levels of P-HER3 protein and NRG1 mRNA were also observed in HER2+ breast cancers progressing after T-DM1 and trastuzumab (NRG1 transcript fold change ± SD; pretreatment = 2, SD = 1.9, vs post-treatment = 11.4, SD = 10.3, P = .04). The HER3-neutralizing antibody LJM716 resensitized the drug-resistant cells to T-DM1, suggesting a causal association between the NRG1-HER3 axis and drug resistance. Finally, Pan-HER treatment inhibited growth of HR6 trastuzumab- and T-DM1-resistant xenografts.Conclusions: These data suggest that upregulation of a NRG1-HER3 axis can mediate escape from anti-HER2 therapies. Further, multitargeted antibody mixtures, such as Pan-HER, can simultaneously remove and/or block targeted ERBB receptor and ligands, thus representing an effective approach against drug-sensitive and -resistant HER2+ cancers. [ABSTRACT FROM AUTHOR]- Published
- 2017
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168. Novel OX40 and 4-1BB derived spacers enhance CD30 CAR activity and safety in CD30 positive lymphoma models.
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Kua L, Ng CH, Tan JW, Tan HC, Seh CC, Wong F, Ong R, Rooney CM, Tan J, Chen Q, Horak ID, Tan KW, and Low L
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- Animals, Humans, Mice, Cell Line, Tumor, Disease Models, Animal, T-Lymphocytes immunology, T-Lymphocytes metabolism, Tumor Necrosis Factor Receptor Superfamily, Member 9 metabolism, Tumor Necrosis Factor Receptor Superfamily, Member 9 immunology, Xenograft Model Antitumor Assays, Immunotherapy, Adoptive methods, Immunotherapy, Adoptive adverse effects, Ki-1 Antigen immunology, Ki-1 Antigen metabolism, Lymphoma therapy, Lymphoma immunology, Receptors, Chimeric Antigen immunology, Receptors, Chimeric Antigen metabolism, Receptors, Chimeric Antigen genetics, Receptors, OX40 metabolism, Receptors, OX40 immunology
- Abstract
The chimeric antigen receptor (CAR) derived from the CD30 specific murine antibody, HRS-3, has produced promising clinical efficacy with a favorable safety profile in the treatment of relapsed or refractory CD30-positive lymphomas. However, persistence of the autologous CAR-T cells was brief, and many patients relapsed a year after treatment. The lack of persistence may be attributed to the use of a wild-type immunoglobulin (Ig)G1 spacer that can associate with Fc receptors. We first identified the cysteine-rich domain (CRD) 5 of CD30 as the primary binding epitope of HRS-3 and armed with this insight, attempted to improve the HRS-3 CAR functionality with a panel of novel spacer designs. We demonstrate that HRS-3 CARs with OX40 and 4-1BB derived spacers exhibited similar anti-tumor efficacy, circumvented interactions with Fc receptors, and secreted lower levels of cytokines in vitro than a CAR employing the IgG1 spacer. Humanization of the HRS-3 scFv coupled with the 4-1BB spacer preserved potent on-target, on-tumor efficacy, and on-target, off-tumor safety. In a lymphoma mouse model of high tumor burden, T cells expressing humanized HRS-3 CD30.CARs with the 4-1BB spacer potently killed tumors with low levels of circulating inflammatory cytokines, providing a promising candidate for future clinical development in the treatment of CD30-positive malignancies., Competing Interests: Declaration of interests L.K., C.H.N., J.W.T., H.C.T., C.C.S., F.W., R.O., I.D.H., K.W.T., and L.L. were employees of Tessa Therapeutics. C.M.R. and Q.C. receive research support from Tessa Therapeutics. This study was funded by Tessa Therapeutics. Patents have been filed based on the work in this study. L.K., J.W.T., F.W., R.O., I.D.H., K.W.T., and L.L. are currently employees of Tikva Allocell., (Copyright © 2024 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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169. Overexpression of an Engineered SERPINB9 Enhances Allogeneic T-cell Persistence and Efficacy.
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Teo PY, Jung Y, Quach DH, Koh J, Ong RW, Goh A, Tan A, Ng CH, Seh CC, Tan KW, Horak ID, and Low L
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- Animals, Humans, Mice, Xenograft Model Antitumor Assays, Cell Line, Tumor, Serpins genetics, Serpins metabolism, Immunotherapy, Adoptive methods, T-Lymphocytes immunology, T-Lymphocytes metabolism, Receptors, Chimeric Antigen immunology, Receptors, Chimeric Antigen genetics, Receptors, Chimeric Antigen metabolism
- Abstract
Allogeneic chimeric antigen receptor (CAR)-expressing T cells offer many advantages over autologous therapies, but their benefits are curtailed by graft-versus-host disease and elimination by recipient immune cells. Moreover, just as with autologous therapies, allogeneic CAR T cells are susceptible to activation-induced cell death (AICD) caused by chronic antigen exposure (CAE). Granzyme B- and Fas/Fas ligand-initiated caspase-mediated apoptoses are key mechanisms of T-cell death caused by T/NK cell-mediated allorejection or CAE. We explored a protective strategy of engineering CAR T cells to overexpress variants of the Granzyme B-specific serine protease inhibitor SERPINB9 (SB9) to improve allogeneic T-cell persistence and antitumor efficacy. We showed that the overexpression of an SB9 variant with broadened caspase specificity, SB9(CAS), not only significantly reduced rejection of allogeneic CAR T cells but also increased their resistance to AICD and enabled them to thrive better under CAE, thus improving allogeneic T-cell persistence and antitumor activity in vitro and in vivo. In addition, although SB9(CAS) overexpression improved the efficacy of allogeneic CAR T-cell therapy by conferring protection to cell death, we did not observe any autonomous growth, and the engineered CAR T cells were still susceptible to an inducible suicide switch. Hence, SB9(CAS) overexpression is a promising strategy that can strengthen current development of cell therapies, broadening their applications to address unmet medical needs., (©2024 American Association for Cancer Research.)
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- 2024
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170. B7-H3-Targeting Chimeric Antigen Receptors Epstein-Barr Virus-specific T Cells Provides a Tumor Agnostic Off-The-Shelf Therapy Against B7-H3-positive Solid Tumors.
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Yeo SP, Kua L, Tan JW, Lim JK, Wong FH, Santos MD, Poh CM, Goh AX, Koh XY, Zhou X, Rajarethinam R, Chen Q, Her Z, Horak ID, Low L, and Tan KW
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- Animals, Humans, Mice, Cell Line, Tumor, Neoplasms therapy, Neoplasms immunology, Female, Single-Domain Antibodies immunology, B7 Antigens immunology, B7 Antigens metabolism, Receptors, Chimeric Antigen immunology, Receptors, Chimeric Antigen metabolism, Herpesvirus 4, Human immunology, Immunotherapy, Adoptive methods, Xenograft Model Antitumor Assays, T-Lymphocytes immunology, T-Lymphocytes metabolism
- Abstract
Encouraged by the observations of significant B7-H3 protein overexpression in many human solid tumors compared to healthy tissues, we directed our focus towards targeting B7-H3 using chimeric antigen receptor (CAR) T cells. We utilized a nanobody as the B7-H3-targeting domain in our CAR construct to circumvent the stability issues associated with single-chain variable fragment-based domains. In efforts to expand patient access to CAR T-cell therapy, we engineered our nanobody-based CAR into human Epstein-Barr virus-specific T cells (EBVST), offering a readily available off-the-shelf treatment. B7H3.CAR-armored EBVSTs demonstrated potent in vitro and in vivo activities against multiple B7-H3-positive human tumor cell lines and patient-derived xenograft models. Murine T cells expressing a murine equivalent of our B7H3.CAR exhibited no life-threatening toxicities in immunocompetent mice bearing syngeneic tumors. Further in vitro evaluation revealed that while human T, B, and natural killer cells were unaffected by B7H3.CAR EBVSTs, monocytes were targeted because of upregulation of B7-H3. Such targeting of myeloid cells, which are key mediators of cytokine release syndrome (CRS), contributed to a low incidence of CRS in humanized mice after B7H3.CAR EBVST treatment. Notably, we showed that B7H3.CAR EBVSTs can target B7-H3-expressing myeloid-derived suppressor cells (MDSC), thereby mitigating MDSC-driven immune suppression. In summary, our data demonstrate that our nanobody-based B7H3.CAR EBVSTs are effective as an off-the-shelf therapy for B7-H3-positive solid tumors. These cells also offer an avenue to modulate the immunosuppressive tumor microenvironment, highlighting their promising clinical potential in targeting solid tumors., Significance: Clinical application of EBVSTs armored with B7-H3-targeting CARs offer an attractive solution to translate off-the-shelf CAR T cells as therapy for solid tumors., (© 2024 The Authors; Published by the American Association for Cancer Research.)
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- 2024
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171. First-in-human trial exploring safety, antitumor activity, and pharmacokinetics of Sym013, a recombinant pan-HER antibody mixture, in advanced epithelial malignancies.
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Berlin J, Tolcher AW, Ding C, Whisenant JG, Horak ID, Wood DL, Nadler PI, Hansen UH, Lantto J, Skartved NJØ, Pedersen MW, and Patnaik A
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- Antibodies, Monoclonal adverse effects, Diarrhea chemically induced, Humans, Maximum Tolerated Dose, Antineoplastic Agents adverse effects, Exanthema chemically induced, Neoplasms metabolism, Neoplasms, Glandular and Epithelial chemically induced, Neoplasms, Glandular and Epithelial drug therapy
- Abstract
Purpose: Sym013 contains six humanized monoclonal antibodies that bind to non-overlapping epitopes on three human epidermal growth factor receptors (HER1-3). Preclinical studies suggested Sym013 strongly suppresses growth of multiple epithelial tumors. This is a first-in-human study exploring safety and efficacy of Sym013 in patients with advanced epithelial malignancies., Methods: Dose escalation used single-patient cohorts until the stopping rule was met, followed by 3 + 3 design. Dose levels planned were: 1, 2, 4, 6, 9, 12, 15, and 18 mg/kg. Treatment cycles were 28 days with imaging every eight weeks. Serum samples were collected at multiple time points for assessment of pharmacokinetics and development of anti-drug antibodies., Results: Thirty-two patients were enrolled with multiple solid tumors, most common being colorectal cancer (CRC; 10/32, 31%). Due to mucositis, rash, and diarrhea at 4 mg/kg once-weekly, dosing was changed to biweekly (Q2W). Mandatory prophylaxis was added due to Grade 3 infusion-related reaction and oral mucositis at 9 mg/kg Q2W. The 15 mg/kg Q2W cohort was enrolling when the study was terminated for business reasons. Most common adverse events were skin (81%) and gastrointestinal (75%) disorders, including dermatitis/rash, stomatitis, and diarrhea. One patient with CRC achieved a partial response; 12 patients with varied malignancies had stable disease., Conclusion: During the conduct of the study, management of frequent infusion-related reactions, skin toxicities, and mucosal disorders, which are indicative of HER inhibition, necessitated multiple protocol amendments. The investigators, in concert with the Sponsor, agreed that achieving a tolerated regimen with acceptable target saturation was unlikely., Trial Registry: www., Clinicaltrials: gov ; NCT02906670 (September 20, 2016)., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
- Published
- 2022
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172. Paediatric Strategy Forum for medicinal product development of chimeric antigen receptor T-cells in children and adolescents with cancer: ACCELERATE in collaboration with the European Medicines Agency with participation of the Food and Drug Administration.
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Pearson AD, Rossig C, Mackall C, Shah NN, Baruchel A, Reaman G, Ricafort R, Heenen D, Bassan A, Berntgen M, Bird N, Bleickardt E, Bouchkouj N, Bross P, Brownstein C, Cohen SB, de Rojas T, Ehrlich L, Fox E, Gottschalk S, Hanssens L, Hawkins DS, Horak ID, Taylor DH, Johnson C, Karres D, Ligas F, Ludwinski D, Mamonkin M, Marshall L, Masouleh BK, Matloub Y, Maude S, McDonough J, Minard-Colin V, Norga K, Nysom K, Pappo A, Pearce L, Pieters R, Pule M, Quintás-Cardama A, Richardson N, Schüßler-Lenz M, Scobie N, Sersch MA, Smith MA, Sterba J, Tasian SK, Weigel B, Weiner SL, Zwaan CM, Lesa G, and Vassal G
- Subjects
- Adolescent, Child, Europe, Humans, Pediatrics, United States, United States Food and Drug Administration, Drug Development organization & administration, Medical Oncology organization & administration, Receptors, Antigen, T-Cell genetics, Receptors, Chimeric Antigen genetics
- Abstract
The seventh multi-stakeholder Paediatric Strategy Forum focused on chimeric antigen receptor (CAR) T-cells for children and adolescents with cancer. The development of CAR T-cells for patients with haematological malignancies, especially B-cell precursor acute lymphoblastic leukaemia (BCP-ALL), has been spectacular. However, currently, there are scientific, clinical and logistical challenges for use of CAR T-cells in BCP-ALL and other paediatric malignancies, particularly in acute myeloid leukaemia (AML), lymphomas and solid tumours. The aims of the Forum were to summarise the current landscape of CAR T-cell therapy development in paediatrics, too identify current challenges and future directions, with consideration of other immune effector modalities and ascertain the best strategies to accelerate their development and availability to children. Although the effect is of limited duration in about half of the patients, anti-CD19 CAR T-cells produce high response rates in relapsed/refractory BCP-ALL and this has highlighted previously unknown mechanisms of relapse. CAR T-cell treatment as first- or second-line therapy could also potentially benefit patients whose disease has high-risk features associated with relapse and failure of conventional therapies. Identifying patients with very early and early relapse in whom CAR T-cell therapy may replace haematopoietic stem cell transplantation and be definitive therapy versus those in whom it provides a more effective bridge to haematopoietic stem cell transplantation is a very high priority. Development of approaches to improve persistence, either by improving T cell fitness or using more humanised/fully humanised products and co-targeting of multiple antigens to prevent antigen escape, could potentially further optimise therapy. Many differences exist between paediatric B-cell non-Hodgkin lymphomas (B-NHL) and BCP-ALL. In view of the very small patient numbers with relapsed lymphoma, careful prioritisation is needed to evaluate CAR T-cells in children with Burkitt lymphoma, primary mediastinal B cell lymphoma and other NHL subtypes. Combination trials of alternative targets to CD19 (CD20 or CD22) should also be explored as a priority to improve efficacy in this population. Development of CD30 CAR T-cell immunotherapy strategies in patients with relapsed/refractory Hodgkin lymphoma will likely be most efficiently accomplished by joint paediatric and adult trials. CAR T-cell approaches are early in development for AML and T-ALL, given the unique challenges of successful immunotherapy actualisation in these diseases. At this time, CD33 and CD123 appear to be the most universal targets in AML and CD7 in T-ALL. The results of ongoing or planned first-in-human studies are required to facilitate further understanding. There are promising early results in solid tumours, particularly with GD2 targeting cell therapies in neuroblastoma and central nervous system gliomas that represent significant unmet clinical needs. Further understanding of biology is critical to success. The comparative benefits of autologous versus allogeneic CAR T-cells, T-cells engineered with T cell receptors T-cells engineered with T cell receptor fusion constructs, CAR Natural Killer (NK)-cell products, bispecific T-cell engager antibodies and antibody-drug conjugates require evaluation in paediatric malignancies. Early and proactive academia and multi-company engagement are mandatory to advance cellular immunotherapies in paediatric oncology. Regulatory advice should be sought very early in the design and preparation of clinical trials of innovative medicines, for which regulatory approval may ultimately be sought. Aligning strategic, scientific, regulatory, health technology and funding requirements from the inception of a clinical trial is especially important as these are very expensive therapies. The model for drug development for cell therapy in paediatric oncology could also involve a 'later stage handoff' to industry after early development in academic hands. Finally, and very importantly, strategies must evolve to ensure appropriate ease of access for children who need and could potentially benefit from these therapies., Competing Interests: Conflict of interest statement The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: ABassan is an employee of Syncopation Life Sciences. EB is an employee of Novartis. CB is an employee of Cellectis. SBC is an employee of CRISPR Therapeutics and has stock ownership in CRISPR. DSH has participated in advisory boards for AstraZeneca and Bayer and has received institutional funding from Incyte, Pfizer, Bristol Myers Squibb, Merck Sharpe Dohme, Lilly. LH is an employee of Miltenyi Biomedicine. IDH is an employee of Tessa Therapeutics. BKM is an employee of Kite, a Gilead company. YM is an employee of Takeda Pharmaceuticals International. SM has participated in advisory boards for Novartis and Wugen and received clinical trial support from Novartis. LP is an employee of GlaxoSmithKline. ADJP has participated in advisory boards for Novartis, Takeda, Merck, Lilly and Celgene and consulted for Lilly and Developmental Therapeutics Consortium Limited MP is an employee of Autolus Limited. AQ-C is an employee of TCR2 Therapeutics. RR is an employee of Celgene/Bristol Myers Squibb. CR has participated in advisory boards for Amgen, BMS, Celgene, Novartis and Pfizer. MAS is an employee and stock ownership of Gracellbiotechnologies Inc. SKT receives research funding from Incyte Corporation and Beam Therapeutics and as participated in advisory boards of Aleta Biotherapeutics and Kura Oncology. MCZ has been a constant for Incyte, Sanofi, BMS, Novartis, Pfizer, Jazz, Abbvie, Roche and Takeda; has received institutional funding from Jazz, Pfizer, Takeda, Abbvie and funding for travel from Jazz. All remaining authors have declared no conflicts of interest., (Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2022
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173. Simultaneous targeting of HER family pro-survival signaling with Pan-HER antibody mixture is highly effective in TNBC: a preclinical trial with PDXs.
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Reddy TP, Choi DS, Anselme AC, Qian W, Chen W, Lantto J, Horak ID, Kragh M, Chang JC, and Rosato RR
- Subjects
- Animals, Cell Proliferation drug effects, Disease Models, Animal, Drug Resistance, Neoplasm, ErbB Receptors antagonists & inhibitors, ErbB Receptors genetics, ErbB Receptors metabolism, Female, Humans, Mice, Molecular Targeted Therapy, Mutation, Receptor, ErbB-2 genetics, Receptor, ErbB-2 metabolism, Receptor, ErbB-3 genetics, Receptor, ErbB-3 metabolism, Triple Negative Breast Neoplasms genetics, Triple Negative Breast Neoplasms metabolism, Triple Negative Breast Neoplasms pathology, Tumor Cells, Cultured, Antibodies, Monoclonal pharmacology, Receptor, ErbB-2 antagonists & inhibitors, Receptor, ErbB-3 antagonists & inhibitors, Triple Negative Breast Neoplasms drug therapy
- Abstract
Background: The human epidermal growth factor receptor (HER) family, notably EGFR, is overexpressed in most triple-negative breast cancer (TNBC) cases and provides cancer cells with compensatory signals that greatly contribute to the survival and development of resistance in response to therapy. This study investigated the effects of Pan-HER (Symphogen, Ballerup, Denmark), a novel mixture of six monoclonal antibodies directed against members of the HER family EGFR, HER2, and HER3, in a preclinical trial of TNBC patient-derived xenografts (PDXs)., Methods: Fifteen low passage TNBC PDX tumor samples were transferred into the right mammary fat pad of mice for engraftment. When tumors reached an average size of 100-200 mm
3 , mice were randomized (n ≥ 6 per group) and treated following three 1-week cycles consisting of three times/week intraperitoneal (IP) injection of either formulation buffer (vehicle control) or Pan-HER (50 mg/kg). At the end of treatment, tumors were collected for Western blot, RNA, and immunohistochemistry analyses., Results: All 15 TNBC PDXs were responsive to Pan-HER treatment, showing significant reductions in tumor growth consistent with Pan-HER-mediated tumor downmodulation of EGFR and HER3 protein levels and significantly decreased activation of associated HER family signaling pathways AKT and ERK. Tumor regression was observed in five of the models, which corresponded to those PDX tumor models with the highest level of HER family activation., Conclusions: The marked effect of Pan-HER in numerous HER family-dependent TNBC PDX models justifies further studies of Pan-HER in TNBC clinical trials as a potential therapeutic option.- Published
- 2020
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174. CD4 + and CD8a + PET imaging predicts response to novel PD-1 checkpoint inhibitor: studies of Sym021 in syngeneic mouse cancer models.
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Kristensen LK, Fröhlich C, Christensen C, Melander MC, Poulsen TT, Galler GR, Lantto J, Horak ID, Kragh M, Nielsen CH, and Kjaer A
- Subjects
- Animals, Antibodies, Monoclonal pharmacology, Antibodies, Neutralizing pharmacology, Biosensing Techniques methods, CD4-Positive T-Lymphocytes metabolism, CD8-Positive T-Lymphocytes metabolism, Cell Line, Tumor, Deferoxamine chemistry, Disease Models, Animal, Immunotherapy, Isografts cytology, Isografts immunology, Lymphocytes, Tumor-Infiltrating immunology, Lymphocytes, Tumor-Infiltrating metabolism, Mice, Molecular Imaging methods, Neoplasms diagnostic imaging, Neoplasms immunology, Programmed Cell Death 1 Receptor immunology, Radioisotopes chemistry, Zirconium chemistry, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Neoplasms drug therapy, Positron-Emission Tomography methods, Programmed Cell Death 1 Receptor antagonists & inhibitors
- Abstract
Predicting the outcome of immunotherapy is essential for efficient treatment. The recent clinical success of immunotherapy is increasingly changing the paradigm of cancer treatment. Accordingly, the development of immune-based agents is accelerating and the number of agents in the global immuno-oncology pipeline has grown 60-70% over the past year. However, despite remarkable clinical efficacy in some patients, only few achieve a lasting clinical response. Treatment failure can be attributed to poorly immunogenic tumors that do not attract tumor infiltrating lymphocytes (TILs). Therefore, we developed positron emission tomography (PET) radiotracers for non-invasive detection of CD4
+ and CD8a+ TILs in syngeneic mouse tumor models for preclinical studies. Methods: Seven syngeneic mouse tumor models (B16F10, P815, CT26, MC38, Renca, 4T1, Sa1N) were quantified for CD4+ and CD8a+ TILs using flow cytometry and immunohistochemistry (IHC), as well as for tumor growth response to Sym021, a humanized PD-1 antibody cross-reactive with mouse PD-1 . Radiotracers were generated from F(ab)'2 fragments of rat-anti-mouse CD4 and CD8a antibodies conjugated to the p -SCN-Bn-Desferrioxamine (SCN-Bn-DFO) chelator and radiolabeled with Zirconium-89 (89 Zr-DFO-CD4/89 Zr-DFO-CD8a). Tracers were optimized for in vivo PET/CT imaging in CT26 tumor-bearing mice and specificity was evaluated by depletion studies and isotype control imaging.89 Zr-DFO-CD4 and89 Zr-DFO-CD8a PET/CT imaging was conducted in the panel of syngeneic mouse models prior to immunotherapy with Sym021. Results: Syngeneic tumor models were characterized as "hot" or "cold" according to number of TILs determined by flow cytometry and IHC.89 Zr-DFO-CD4 and89 Zr-DFO-CD8a were successfully generated with a radiochemical purity >99% and immunoreactivity >85%. The optimal imaging time-point was 24 hours post-injection of ~1 MBq tracer with 30 µg non-labeled co-dose. Reduced tumor and spleen uptake of89 Zr-DFO-CD8a was observed in CD8a+ depleted mice and the uptake was comparable with that of isotype control (89 Zr-DFO-IgG2b) confirming specificity. PET imaging in syngeneic tumor models revealed a varying maximum tumor-to-heart ratio of89 Zr-DFO-CD4 and89 Zr-DFO-CD8a across tumor types and in-between subjects that correlated with individual response to Sym021 at day 10 relative to start of therapy ( p=0.0002 and p=0.0354 , respectively). The maximum89 Zr-DFO-CD4 tumor-to-heart ratio could be used to stratify mice according to Sym021 therapy response and overall survival was improved in mice with a89 Zr-DFO-CD4 ratio >9 ( p=0.0018 ). Conclusion: We developed89 Zr-DFO-CD4 and89 Zr-DFO-CD8a PET radiotracers for specific detection and whole-body assessment of CD4+ and CD8a+ status. These radiotracers can be used to phenotype preclinical syngeneic mouse tumor models and to predict response to an immune checkpoint inhibitor. We foresee development of such non-invasive in vivo biomarkers for prediction and evaluation of clinical efficacy of immunotherapeutic agents, such as Sym021., Competing Interests: Competing Interests: C.F., M.C.M., T.T.P., G.R.G, J.L., I.D.H. and M.K. are employed by Symphogen A/S that holds the right of Sym021. All other authors declare no competing interests., (© The author(s).)- Published
- 2019
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175. Sym004-induced EGFR elimination is associated with profound anti-tumor activity in EGFRvIII patient-derived glioblastoma models.
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Keir ST, Chandramohan V, Hemphill CD, Grandal MM, Melander MC, Pedersen MW, Horak ID, Kragh M, Desjardins A, Friedman HS, and Bigner DD
- Subjects
- Animals, Brain Neoplasms genetics, Brain Neoplasms metabolism, Brain Neoplasms pathology, ErbB Receptors genetics, ErbB Receptors immunology, ErbB Receptors metabolism, Female, Glioblastoma genetics, Glioblastoma metabolism, Glioblastoma pathology, Humans, Male, Mice, Inbred BALB C, Mice, Nude, Subcutaneous Tissue, Xenograft Model Antitumor Assays, Antibodies, Monoclonal administration & dosage, Antineoplastic Agents administration & dosage, Brain Neoplasms therapy, Glioblastoma therapy
- Abstract
Background: Sym004 is a mixture of two monoclonal antibodies (mAbs), futuximab and modotuximab, targeting non-overlapping epitopes on the epidermal growth factor receptor (EGFR). Previous studies have shown that Sym004 is more efficient at inducing internalization and degradation of EGFR than individual components, which translates into superior cancer cell inhibition. We investigated whether Sym004 induces removal of EGFRvIII and if this removal translates into tumor growth inhibition in hard-to-treat glioblastomas (GBMs) harboring the mutated, constitutively active EGFR variant III (EGFRvIII)., Methods: To address this question, we tested the effect of Sym004 versus cetuximab in eight patient-derived GBM xenograft models expressing either wild-type EGFR (EGFRwt) and/or mutant EGFRvIII. All models were tested as both subcutaneous and orthotopic intracranial xenograft models., Results: In vitro studies demonstrated that Sym004 internalized and removed EGFRvIII more efficiently than mAbs, futuximab, modotuximab, and cetuximab. Removal of EGFRvIII by Sym004 translated into significant in vivo anti-tumor activity in all six EGFRvIII xenograft models. Furthermore, the anti-tumor activity of Sym004 in vivo was superior to that of its individual components, futuximab and modotuximab, suggesting a clear synergistic effect of the mAbs in the mixture., Conclusion: These results demonstrate the broad activity of Sym004 in patient-derived EGFRvIII-expressing GBM xenograft models and provide a clear rationale for clinical evaluation of Sym004 in EGFRvIII-positive adult GBM patients.
- Published
- 2018
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176. Pan-HER-An antibody mixture targeting EGFR, HER2 and HER3 abrogates preformed and ligand-induced EGFR homo- and heterodimers.
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Ellebaek S, Brix S, Grandal M, Lantto J, Horak ID, Kragh M, and Poulsen TT
- Subjects
- Cell Line, Tumor, Cell Proliferation drug effects, Dimerization, Drug Resistance, Neoplasm drug effects, ErbB Receptors metabolism, Gene Expression Regulation, Neoplastic, Humans, Neoplasms chemistry, Neoplasms drug therapy, Protein Binding drug effects, Receptor, ErbB-2 metabolism, Receptor, ErbB-3 metabolism, Antibodies, Monoclonal, Humanized pharmacology, ErbB Receptors chemistry, Neoplasms metabolism, Receptor, ErbB-2 chemistry, Receptor, ErbB-3 chemistry
- Abstract
The human epidermal growth factor receptor (HER)-family is involved in development of many epithelial cancers. Therefore, HER-family members constitute important targets for anti-cancer therapeutics such as monoclonal antibodies (mAbs). A limitation to the success of single HER-targeting mAbs is development of acquired resistance through mechanisms such as alterted receptor dimerization patterns and dependencies. Pan-HER is a mixture of six mAbs simultaneously targeting epidermal growth factor receptor (EGFR), HER2 and HER3 with two mAbs against each receptor. Pan-HER has previously demonstrated broader efficacy than targeting single or dual receptor combinations also in resistant settings. In light of this broad efficacy, we decided to investigate the effect of Pan-HER compared with single HER-targeting with single and dual mAbs on HER-family cross-talk and dimerization focusing on EGFR. The effect of Pan-HER on cell proliferation and HER-family receptor degradation was superior to treatment with single mAbs targeting either single receptor, and similar to targeting a single receptor with two non-overlapping antibodies. Furthermore, changes in EGFR-dimerization patterns after treatment with Pan-HER were investigated by in situ proximity ligation assay and co-immunoprecipitation, demonstrating that Pan-HER and the EGFR-targeting mAb mixture efficiently down-regulate basal EGFR homo- and heterodimerization in two tested cell lines, whereas single mAbs had limited effects. Pan-HER and the EGFR-targeting mAb mixture also blocked EGF-binding and thereby ligand-induced changes in EGFR-dimerization levels. These results suggest that Pan-HER reduces the cellular capability to switch HER-dependency and dimerization pattern in response to treatment and thus hold promise for future clinical development of Pan-HER in resistant settings., (© 2016 UICC.)
- Published
- 2016
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177. In vivo imaging of therapy response to a novel pan-HER antibody mixture using FDG and FLT positron emission tomography.
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Nielsen CH, Jensen MM, Kristensen LK, Dahlman A, Fröhlich C, Jacobsen HJ, Poulsen TT, Lantto J, Horak ID, Kragh M, and Kjaer A
- Subjects
- Adenocarcinoma drug therapy, Adenocarcinoma genetics, Adenocarcinoma immunology, Animals, Cell Line, Tumor, Cell Proliferation drug effects, ErbB Receptors immunology, Female, Gene Expression Regulation, Neoplastic drug effects, Humans, Mice, Nude, Multimodal Imaging, Pancreatic Neoplasms genetics, Pancreatic Neoplasms immunology, Predictive Value of Tests, Receptor, ErbB-2 immunology, Receptor, ErbB-3 immunology, Time Factors, Tumor Burden drug effects, X-Ray Microtomography, Xenograft Model Antitumor Assays, Adenocarcinoma diagnostic imaging, Antibodies, Monoclonal pharmacology, Antineoplastic Combined Chemotherapy Protocols pharmacology, Dideoxynucleosides administration & dosage, ErbB Receptors antagonists & inhibitors, Fluorodeoxyglucose F18 administration & dosage, Pancreatic Neoplasms diagnostic imaging, Pancreatic Neoplasms drug therapy, Positron-Emission Tomography methods, Radiopharmaceuticals administration & dosage, Receptor, ErbB-2 antagonists & inhibitors, Receptor, ErbB-3 antagonists & inhibitors
- Abstract
Purpose: Overexpression of the human epidermal growth factor receptor (HER) family and their ligands plays an important role in many cancers. Targeting multiple members of the HER family simultaneously may increase the therapeutic efficacy. Here, we report the ability to image the therapeutic response obtained by targeting HER family members individually or simultaneously using the novel monoclonal antibody (mAb) mixture Pan-HER., Experimental Design and Results: Mice with subcutaneous BxPC-3 pancreatic adenocarcinomas were divided into five groups receiving vehicle or mAb mixtures directed against either EGFR (HER1), HER2, HER3 or all three receptors combined by Pan-HER. Small animal positron emission tomography/computed tomography (PET/CT) with 2'-deoxy-2'-[(18)F]fluoro-D-glucose (FDG) and 3'-deoxy-3'-[(18)F]fluorothymidine (FLT) was performed at baseline and at day 1 or 2 after initiation of therapy. Changes in tumor uptake of tracers were quantified and compared to reduction in tumor size. Imaging results were further validated by immunohistochemistry and qPCR. Mean FDG and FLT uptake in the Pan-HER treated group decreased by 19 ± 4.3% and 24 ± 3.1%, respectively. The early change in FDG and FLT uptake correlated with tumor growth at day 23 relative to day 0. Ex vivo molecular analyses of markers associated with the mechanisms of FDG and FLT uptake confirmed the in vivo imaging results., Conclusions: Taken together, the study supports the use of FDG and FLT as imaging biomarkers of early response to Pan-HER therapy. FDG and FLT PET/CT imaging should be considered as imaging biomarkers in clinical evaluation of the Pan-HER mAb mixture.
- Published
- 2015
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178. Potent and sustained inhibition of HIF-1α and downstream genes by a polyethyleneglycol-SN38 conjugate, EZN-2208, results in anti-angiogenic effects.
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Sapra P, Kraft P, Pastorino F, Ribatti D, Dumble M, Mehlig M, Wang M, Ponzoni M, Greenberger LM, and Horak ID
- Subjects
- Animals, Antineoplastic Agents, Phytogenic pharmacology, Camptothecin pharmacology, Cell Line, Tumor, Glioma metabolism, Glioma pathology, Humans, Irinotecan, Mice, Mice, Nude, Neovascularization, Pathologic metabolism, Neovascularization, Pathologic pathology, Xenograft Model Antitumor Assays, Angiogenesis Inhibitors pharmacology, Camptothecin analogs & derivatives, Down-Regulation drug effects, Gene Expression Regulation, Neoplastic drug effects, Glioma drug therapy, Hypoxia-Inducible Factor 1, alpha Subunit biosynthesis, Neoplasm Proteins biosynthesis, Neovascularization, Pathologic drug therapy, Polyethylene Glycols pharmacology
- Abstract
Topoisomerase I inhibitors down-regulate HIF-1α leading to tumor growth inhibition, but only while maintaining sustained levels of drug exposure. EZN-2208, a multi-arm 40 kDa pegylated, releasable SN38-drug conjugate, provides higher, longer lasting exposure of tumors to SN38 in contrast to SN38 that is released from CPT-11. EZN-2208 also consistently has greater antitumor activity than CPT-11 in a variety of solid and hematological tumor models. In this report, the ability of PEG-SN38 to down-regulate HIF-1α and its downstream targets, in a more potent, sustained manner compared with CPT-11 was examined. To do so, U251 glioma xenografts that stably expressed a hypoxia response element-dependent luciferase reporter gene were implanted in mice. After treatment it was found that EZN-2208 induced potent, sustained HIF-1α down-regulation (37% at 48 h and 83% at 120 h) in the tumors, whereas CPT-11 caused only minor, transient HIF-1α down-regulation. In addition, EZN-2208 down-regulated mRNA levels of HIF-1α targeted genes (MMP2, VEGF1, Glut1, Glut3 and TGFβ1). Further, western blot analyses of xenograft tumors demonstrated that EZN-2208 had significantly more effect than CPT-11 in down-regulating HIF-1α, VEGF, Glut1 and MMP2 protein levels. Significant down-regulation of HIF-1α and VEGF proteins translated to EZN-2208's superior anti-angiogenic activity compared with CPT-11, confirmed by microvessel density reduction in a chorioallantoic membrane assay and in CD-31 immunohistochemistry studies. Additional studies done with matrigel implants devoid of tumor cells show that EZN-2208 significantly inhibits angiogenesis while CPT-11 has little or no effect. It is concluded that the superior antitumor activity of EZN-2208 compared with CPT-11 is attributed, in part, to an anti-angiogenic effect. Ongoing clinical Phase I and Phase II studies will assess safety and efficacy of EZN-2208.
- Published
- 2011
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179. Marked therapeutic efficacy of a novel polyethylene glycol-SN38 conjugate, EZN-2208, in xenograft models of B-cell non-Hodgkin's lymphoma.
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Sapra P, Kraft P, Mehlig M, Malaby J, Zhao H, Greenberger LM, and Horak ID
- Subjects
- Animals, Burkitt Lymphoma drug therapy, Burkitt Lymphoma pathology, Camptothecin chemistry, Camptothecin therapeutic use, Cell Line, Tumor, Female, Humans, Lymphoma, B-Cell pathology, Mice, Mice, SCID, Camptothecin analogs & derivatives, Lymphoma, B-Cell drug therapy, Polyethylene Glycols chemistry, Polyethylene Glycols therapeutic use, Xenograft Model Antitumor Assays methods
- Abstract
Examination of the clinical utility of SN38 (10-hydroxy-7-ethyl-camptothecin), the active metabolite of CPT-11, has not been possible to date due to poor solubility of SN38. Here we evaluated the activity of EZN-2208, a water-soluble polyethyleneglycol-SN38 conjugate, in pre-clinical models of Burkitt's non-Hodgkin's lymphoma (NHL) (Raji and Daudi), and follicular NHL (DoHH2). In vitro, the IC50 of EZN-2208 ranged from 3-24 nM, which was 30- to 45-fold lower than CPT-11 or 2.5- to 3.5-fold higher than SN38. In both an early-disease Raji model and an advanced-disease Daudi model, treatment with multiple doses of EZN-2208 resulted in 90% and 100% cures of animals, respectively (cure defined as no sign of tumors by gross observations at the termination of study). The activity of EZN-2208 was dramatically superior to that of CPT-11 in all three models. The excellent therapeutic efficacy of EZN-2208 in several B-cell NHL xenograft models merits its evaluation in the clinic for lymphoid malignancies.
- Published
- 2009
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180. Clinical-scale radiolabeling of a humanized anticarcinoembryonic antigen monoclonal antibody, hMN-14, with residualizing 131I for use in radioimmunotherapy.
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Govindan SV, Griffiths GL, Stein R, Andrews P, Sharkey RM, Hansen HJ, Horak ID, and Goldenberg DM
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- Antibodies, Monoclonal isolation & purification, Antibodies, Monoclonal, Humanized, Carcinoembryonic Antigen isolation & purification, Drug Stability, Humans, Oligopeptides chemical synthesis, Oligopeptides isolation & purification, Pentetic Acid chemical synthesis, Pentetic Acid chemistry, Pentetic Acid isolation & purification, Pentetic Acid therapeutic use, Radiopharmaceuticals chemical synthesis, Radiopharmaceuticals isolation & purification, Antibodies, Monoclonal chemistry, Antibodies, Monoclonal therapeutic use, Carcinoembryonic Antigen chemistry, Carcinoembryonic Antigen immunology, Isotope Labeling methods, Oligopeptides chemistry, Oligopeptides therapeutic use, Pentetic Acid analogs & derivatives, Radioimmunotherapy methods, Radiopharmaceuticals chemistry, Radiopharmaceuticals therapeutic use
- Abstract
Unlabelled: Radiolabeling of monoclonal antibodies (mAbs) with an intracellularly trapped form of (131)I (residualizing (131)I) involves radioiodinating a small molecular entity, conjugating it to the mAb, and purification. Column purifications are impractical during procedures involving multi-gigabecquerel levels of radioactivity. The goal of this study was to develop a simple, remote, "1-pot" method of radiolabeling and purification for the scaled-up radioiodination of a humanized anti-carcinoembryonic antigen (CEA) mAb, humanized MN-14 (hMN-14; labetuzumab), with an optimized residualizing (131)I moiety, (131)I-IMP-R4. IMP-R4 is MCC-Lys(MCC)-Lys(X)-d-Tyr-d-Lys(X)-OH, where MCC is 4-(N-maleimidomethyl)-cyclohexane-1-carbonyl and X is 1-((4-thiocarbonylamino)benzyl)-diethylenetriaminepentaacetic acid., Methods: An IODO-GEN-based remote labeling system was used. IMP-R4 was radioiodinated (0.13 mumol per 3.7 GBq of (131)I) at a pH of 7.0-7.4 and conjugated to disulfide-reduced hMN-14 after quenching of unused reactive (131)I. The product was purified by stirring for 5 min with a 20% (w/v) suspension of an anion-exchange resin and sterilely filtered into a sealed vial. Human serum albumin was added at a final concentration of 1%-2.5%. Immunoreactivity was determined by mixing with CEA and determining the complexation level by size-exclusion high-pressure liquid chromatography. Two control radiolabelings, either with unreduced hMN-14 or with IMP-R4 omitted, also were performed., Results: In 18 radiolabelings with (131)I in the range of 2.04-4.81 GBq (55-130 mCi), yields of 59.9% +/- 7.9% (mean +/- SD) at specific activities of 200 +/- 26 MBq/mg (5.4 +/- 0.7 mCi/mg) were obtained, with > or =95% of the radioactivity being associated with hMN-14 and with < or =4% aggregation. Similar yields were obtained in a subset of radiolabelings (n = 7) with >3.7 GBq of (131)I. The immunoreactivities of the preparations were typically >95%. Nonspecific incorporation in the absence of IMP-R4 was 0.5%, whereas that obtained with unreduced IgG was approximately 8%, possibly because of conjugation of IMP-R4 at lysine sites. The process also removed >99% of the quenching reagent used. Radiolabelings performed with freshly prepared solutions or lyophilized preparations produced similar yields, a result that suggested the option for a single-use kit design., Conclusion: Efficient removal of (131)I-IMP-R4 and quenched (131)I by 5 min of stirring with anion-exchange resin renders a multi-gigabecquerel-level preparation of (131)I-IMP-R4-hMN-14 safe, convenient, and practical.
- Published
- 2005
181. Monoclonal Antibodies in Cancer--Fourth International Congress. 3-6 September 2004, Colorado Springs, CO, USA.
- Author
-
Horak ID
- Subjects
- Animals, Humans, Antibodies, Monoclonal pharmacology, Antineoplastic Agents pharmacology
- Published
- 2004
182. Reagents and methods for PET using bispecific antibody pretargeting and 68Ga-radiolabeled bivalent hapten-peptide-chelate conjugates.
- Author
-
Griffiths GL, Chang CH, McBride WJ, Rossi EA, Sheerin A, Tejada GR, Karacay H, Sharkey RM, Horak ID, Hansen HJ, and Goldenberg DM
- Subjects
- Animals, Cell Line, Tumor, Chelating Agents chemistry, Haptens chemistry, Humans, Indicators and Reagents chemistry, Metabolic Clearance Rate, Mice, Mice, Nude, Oligopeptides chemistry, Organ Specificity, Organometallic Compounds chemistry, Radiopharmaceuticals pharmacokinetics, Tissue Distribution, Antibodies, Bispecific chemistry, Colonic Neoplasms diagnostic imaging, Colonic Neoplasms metabolism, Isotope Labeling methods, Oligopeptides pharmacokinetics, Organometallic Compounds pharmacokinetics, Tomography, Emission-Computed methods
- Abstract
Unlabelled: The aim of this work was to develop reagents and methods potentially useful in PET, using (68)Ga in a 2-step pretargeting protocol., Methods: We prepared bispecific antibodies (bsAbs) for disease-specific targeting of carcinoembryonic antigen-positive cells and recognition of later-administered bivalent hapten-peptide conjugates. The secondary antibody arm (antibody 679) recognizes a histaminyl-succinyl-glycine (HSG) structural subunit. The bsAbs were prepared as Fab' x Fab' conjugates using chemical cross-linking methods and as bispecific diabodies using recombinant DNA technologies. A HSG-bivalent hapten conjugate bearing the macrocyclic ring chelating agent 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid (DOTA) was designed to be readily radiolabeled with (68)Ga taken directly from a (68)Ge/(68)Ga generator system. Reagents were tested in vitro and, then, for their targeting properties in a preclinical animal model of human cancer., Results: A chemically cross-linked hMN-14 x 679 F(ab')(2) and a fully humanized bispecific diabody construct (BS1.5H), expressed in Escherichia coli, were prepared for this work. We synthesized the bivalent peptide termed IMP 241 [DOTA-Phe-Lys(HSG)-D-Tyr-Lys(HSG)-NH(2)] and labeled it with (68)Ga and (67)Ga at temperatures from 45 degrees C to 100 degrees C, over times of 15 min to 1 h, establishing 15 min at 95 degrees C as a useful condition for (68)Ga labeling. When we formulated the IMP 241 bivalent hapten-peptide with ammonium acetate buffer at pH 4-5 and eluted the (68)Ga from the generator directly into the peptide solution, we achieved an almost quantitative incorporation of the (68)Ga into IMP 241, as analyzed by size-exclusion high-performance liquid chromatography, after mixing the complex with the 679 antibody. For in vivo studies we used (67)Ga-IMP 241 as a surrogate for (68)Ga-IMP 241, in view of the short, 68-min half-life of the (68)Ga nuclide. The (67)Ga-IMP 241 was successfully pretargeted to human colon tumor xenografts in athymic mice with both the chemical and the diabody bispecific proteins. High tumor-to-normal tissue ratios for (67)Ga uptake were found for all tissues at 1 to 6 h after injection of (67)Ga-IMP 241. When using the BS1.5H diabody for pretargeting, tumor-to-blood, tumor-to-liver, and tumor-to-lung ratios of (67)Ga-IMP 241 at 1 and 3 h after injection were 41:1 and 137:1, 51:1 and 106:1, and 16:1 and 46:1, respectively., Conclusion: The general approach described, along with the new compositions and the labeling methods we have developed, may eventually allow for use of (68)Ga-labeled specific targeting agents in a routine clinical PET application.
- Published
- 2004
183. Cure of SCID mice bearing human B-lymphoma xenografts by an anti-CD74 antibody-anthracycline drug conjugate.
- Author
-
Griffiths GL, Mattes MJ, Stein R, Govindan SV, Horak ID, Hansen HJ, and Goldenberg DM
- Subjects
- Animals, Antibiotics, Antineoplastic therapeutic use, Antibodies, Monoclonal chemistry, Antigens, Differentiation, B-Lymphocyte biosynthesis, Disulfides chemistry, Doxorubicin pharmacology, Histocompatibility Antigens Class II biosynthesis, Humans, Immunohistochemistry, Immunotherapy, Mice, Mice, SCID, Models, Chemical, Neoplasm Transplantation, Neoplasms, Experimental therapy, Time Factors, Anthracyclines pharmacology, Antigens, Differentiation, B-Lymphocyte chemistry, Histocompatibility Antigens Class II chemistry, Lymphoma, B-Cell immunology
- Abstract
Purpose: The purpose of this research was to test the therapeutic efficacy of an anthracycline-antibody conjugate for the treatment of human B-cell lymphoma in a preclinical animal model., Experimental Design: Doxorubicin (dox) conjugates of the murine and humanized versions of the anti B-cell antibody LL1, targeting CD74, were prepared, along with a nonspecific control dox-antibody conjugate, targeting carcinoembryonic antigen. Antibody conjugates carried approximately 8-10 drug molecules attached site-specifically at thiols of reduced interchain disulfide bonds. Conjugates were tested, initially in vitro, and then for therapeutic efficacy in a systemic model, using a lethal i.v. dose of Raji cells in SCID mice., Results: Dox-LL1 conjugates were shown to be stable and 3-fold more effective in vitro against the human B-cell Burkitt's lymphoma line, Raji, compared with the nonspecific control conjugate that did not target CD74 or B cells. When SCID mice were given an i.v. dose of 2.5 million Raji cells, they would die of disseminated disease within 15-25 days postinjection. A single dose of dox-LL1 conjugate, 117-350 micro g, given 5 days to 14 (advanced disease) days after injection of the Raji cells resulted in cure of most animals out to 180 days after injection of the cells, whereas animals in treatment control groups were not cured. The dose of dox-LL1 found useful in this work corresponds with a significantly lower drug dose than reported previously with other drug-antibody conjugates, Conclusion: CD74 appears to be a uniquely useful target antigen for delivery of drugs, effecting cures of animals with single, low doses of conjugate.
- Published
- 2003
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