Impact of charge patches on tumor disposition and biodistribution of therapeutic antibodies

This study explores the impact of antibody surface charge on tissue distribution into various tissues including tumor. Tumor-bearing mice were dosed intravenously with a mixture of three antibodies engineered to carry negative charge patches, a balanced charge distribution, or positive patches, respectively. Tissue levels were analyzed with a specific LC-MS/MS method. In addition, the antibody mix was administered to non-tumor bearing mice. Muscle and skin interstitial fluid were obtained by centrifugation and analyzed by LC-MS/MS. An in-vitro endothelium model was explored for its feasibility to mimic the observed distribution differences. A balanced charge distribution was optimal in terms of total tumor exposure, while in other tissues negatively charged and balanced charged antibodies gave similar results. In contrast, positive charge patches generally result in increased serum clearance but markedly enhance tumor and organ uptake, leading to higher tissue-to-serum ratios. The uptake and availability in the interstitial space were confirmed by specific assessment of antibody levels in the interstitial fluid of muscle and skin, with similar charge impact as in total tissue. The in vitro model was able to differentiate the transport propensity of this series of antibody variants. In summary, our results show the differential effects of charge patches on an antibody surface on biodistribution and tumor uptake. These insights may help in the design of molecules with biodistribution properties tailored to their purpose and an optimized safety profile.

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CAFs affect the proliferation and treatment response of head and neck cancer spheroids during co-culturing in a unique in vitro model

Cancer Cell International ◽

10.1186/s12935-020-01718-6 ◽

2020 ◽

Vol 20 (1) ◽

Author(s):

Mustafa Magan ◽

Emilia Wiechec ◽

Karin Roberg

Keyword(s):

Cell Proliferation ◽

Tumor Cell ◽

Head And Neck ◽

Treatment Response ◽

Tumor Cells ◽

In Vitro Model ◽

Tumor Spheroids ◽

Vitro Model ◽

The Impact

Abstract Background Head and neck squamous cell carcinoma (HNSCC) is a heterogeneous group of tumors for which the overall survival rate worldwide is around 60%. The tumor microenvironment, including cancer-associated fibroblasts (CAFs), is believed to affect the treatment response and migration of HNSCC. The aim of this study was to create a biologically relevant HNSCC in vitro model consisting of both tumor cells and CAFs cultured in 3D to establish predictive biomarkers for treatment response, as well as to investigate the impact of CAFs on phenotype, proliferation and treatment response in HNSCC cells. Methods Three different HNSCC patient-derived tumor cell lines were cultured with and without CAFs in a 3D model. Immunohistochemistry of the proliferation marker Ki67, epidermal growth factor receptor (EGFR) and fibronectin and a TUNEL-assay were performed to analyze the effect of CAFs on both tumor cell proliferation and response to cisplatin and cetuximab treatment in tumor spheroids (3D). mRNA expression of epithelial-mesenchymal transition (EMT) and cancer stem cells markers were analyzed using qRT-PCR. Results The results demonstrated increased cell proliferation within the tumor spheroids in the presence of CAFs, correlating with increased expression of EGFR. In spheroids with increased expression of EGFR, a potentiated response to cetuximab treatment was observed. Surprisingly, an increase in Ki67 expressing tumor cells were observed in spheroids treated with cisplatin for 3 days, correlating with increased expression of EGFR. Furthermore, tumor cells co-cultured with CAFs presented an increased EMT phenotype compared to tumor cells cultured alone in 3D. Conclusion Taken together, our results reveal increased cell proliferation and elevated expression of EGFR in HNSCC tumor spheroids in the presence of CAFs. These results, together with the altered EMT phenotype, may influence the response to cetuximab or cisplatin treatment.

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Impact of Bacterial Biofilm Formation on In Vitro and In Vivo Activities of Antibiotics

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.42.4.895 ◽

1998 ◽

Vol 42 (4) ◽

pp. 895-898 ◽

Cited By ~ 112

Author(s):

Silvia Schwank ◽

Zarko Rajacic ◽

Werner Zimmerli ◽

Jürg Blaser

Keyword(s):

Animal Model ◽

Staphylococcus Epidermidis ◽

Biofilm Formation ◽

Bacterial Biofilm ◽

Phenotypic Resistance ◽

Vitro Model ◽

The Impact ◽

Isogenic Strains

ABSTRACT The impact of bacterial adherence on antibiotic activity was analyzed with two isogenic strains of Staphylococcus epidermidis that differ in the features of their in vitro biofilm formation. The eradication of bacteria adhering to glass beads by amikacin, levofloxacin, rifampin, or teicoplanin was studied in an animal model and in a pharmacokinetically matched in vitro model. The features of S. epidermidis RP62A that allowed it to grow on surfaces in multiple layers promoted phenotypic resistance to antibiotic treatment, whereas strain M7 failed to accumulate, despite initial adherence on surfaces and growth in suspension similar to those for RP62A. Biofilms of S. epidermidis M7 were better eradicated than those of strain RP62A in vitro (46 versus 31%;P < 0.05) as well as in the animal model (39 versus 9%; P < 0.01).

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Influence of Bacillus subtilis C-3102 on microbiota in a dynamic in vitro model of the gastrointestinal tract simulating human conditions

Beneficial Microbes ◽

10.3920/bm2012.0016 ◽

2012 ◽

Vol 3 (3) ◽

pp. 229-236 ◽

Cited By ~ 21

Author(s):

M. Hatanaka ◽

Y. Nakamura ◽

A.J.H. Maathuis ◽

K. Venema ◽

I. Murota ◽

...

Keyword(s):

Bacillus Subtilis ◽

Gastrointestinal Tract ◽

In Vitro Model ◽

Germination Rate ◽

Human Colon ◽

Micro Array ◽

Vitro Model ◽

Oral Doses ◽

The Impact

Survival and germination rate of Bacillus subtilis C-3102 spores were investigated in a stomach and small intestine model (TIM-1), while the impact of C-3102 cells that had passed through TIM-1 on human colon microbiota was evaluated in a model of the large intestine (TIM-2). The survival of C-3102 spores in TIM-1 was 99%; 8% of the spores had germinated. Effluent of TIM-1 was subsequently introduced into TIM-2 and a micro-array platform was employed to assess changes in the microbiota composition. The effluent, which contained germinated C-3102 cells, increased some Bifidobacterium species and decreased some Clostridium groups. These changes were greater compared to those obtained by adding C-3102 spores directly to TIM-2. The present study suggests that oral doses of B. subtilis C-3102 spores have the potential to modulate the human colon microbiota. This effect may be caused by germination of the spores in the gastrointestinal tract.

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Digital twinning of Cellular Capsule Technology: Emerging outcomes from the perspective of porous media mechanics

PLoS ONE ◽

10.1371/journal.pone.0254512 ◽

2021 ◽

Vol 16 (7) ◽

pp. e0254512

Author(s):

Stéphane Urcun ◽

Pierre-Yves Rohan ◽

Wafa Skalli ◽

Pierre Nassoy ◽

Stéphane P. A. Bordas ◽

...

Keyword(s):

Tumor Cells ◽

Mechanical Model ◽

Interstitial Fluid ◽

Sensitivity Analyses ◽

Mechanical Forces ◽

Alginate Capsules ◽

Phenotype Switch ◽

New Perspective ◽

The Impact

Spheroids encapsulated within alginate capsules are emerging as suitable in vitro tools to investigate the impact of mechanical forces on tumor growth since the internal tumor pressure can be retrieved from the deformation of the capsule. Here we focus on the particular case of Cellular Capsule Technology (CCT). We show in this contribution that a modeling approach accounting for the triphasic nature of the spheroid (extracellular matrix, tumor cells and interstitial fluid) offers a new perspective of analysis revealing that the pressure retrieved experimentally cannot be interpreted as a direct picture of the pressure sustained by the tumor cells and, as such, cannot therefore be used to quantify the critical pressure which induces stress-induced phenotype switch in tumor cells. The proposed multiphase reactive poro-mechanical model was cross-validated. Parameter sensitivity analyses on the digital twin revealed that the main parameters determining the encapsulated growth configuration are different from those driving growth in free condition, confirming that radically different phenomena are at play. Results reported in this contribution support the idea that multiphase reactive poro-mechanics is an exceptional theoretical framework to attain an in-depth understanding of CCT experiments, to confirm their hypotheses and to further improve their design.

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The role of Visfatin and Resistin in an In Vitro Model of Obesity-Induced Invasive Liver Cancer

Canadian Journal of Physiology and Pharmacology ◽

10.1139/cjpp-2020-0377 ◽

2020 ◽

Author(s):

Candace Miethe ◽

Linda Torres ◽

Jessica Beristain ◽

Megan Zamora ◽

Ramona Salcedo Price

Keyword(s):

Liver Cancer ◽

Cancer Progression ◽

Neutralizing Antibodies ◽

Normal Weight ◽

Cancer Phenotype ◽

Vitro Model ◽

The Impact ◽

Metalloproteinase 9

Background: Obesity is associated with the development of liver disease and its progression to hepatocellular carcinoma. This link may be attributed to adipocytokines such as growth visfatin, and resistin which have been shown to promote liver cancer incidence and progression. Studies have yet to determine the role of visfatin and resistin in liver cancer specifically in the context of obesity. The objective of this study was to investigate the effect of neutralizing visfatin and resistin in obese (OB) or normal weight (NW) sera to determine the contribution of these proteins in obesity-induced invasive liver cancer. Methods: Sera from OB or NW males was used to determine the efficacy of neutralizing visfatin and resistin to reduce the obesity-induced liver cancer phenotype. HepG2 and SNU-449 cells were exposed to OB and NW sera ± antibodies for visfatin or resistin. The neutralizing antibodies differentially suppressed invasion, ROS production, and matrix metalloproteinase-9 secretion. These changes corresponded with a decrease in phosphorylated ERK and Akt in HepG2 cells, but differences were not observed in CAP1 or β-catenin. In conclusion, visfatin and resistin have differential roles in obesity-associated liver cancer and may be potential targets to reverse the impact of obesity on liver cancer progression.

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An in vitro model to evaluate the impact of environmental fine particles (PM0.3-2.5) on skin damage

Toxicology Letters ◽

10.1016/j.toxlet.2019.01.016 ◽

2019 ◽

Vol 305 ◽

pp. 94-102 ◽

Cited By ~ 7

Author(s):

Anthony Verdin ◽

Fabrice Cazier ◽

Richard Fitoussi ◽

Natacha Blanchet ◽

Katell Vié ◽

...

Keyword(s):

In Vitro Model ◽

Fine Particles ◽

Skin Damage ◽

Vitro Model ◽

The Impact

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Intratumoral injection reduces toxicity and antibody-mediated neutralization of immunocytokine in a mouse melanoma model

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-001262 ◽

2020 ◽

Vol 8 (2) ◽

pp. e001262

Author(s):

Claire C Baniel ◽

Elizabeth G Sumiec ◽

Jacqueline A Hank ◽

Amber M Bates ◽

Amy K Erbe ◽

...

Keyword(s):

Tumor Regression ◽

Interleukin 2 ◽

Intratumoral Injection ◽

In Vitro Assays ◽

Human Antibody ◽

Severe Toxicity ◽

Tumor Bearing ◽

The Impact

BackgroundSome patients with cancer treated with anticancer monoclonal antibodies (mAbs) develop antidrug antibodies (ADAs) that recognize and bind the therapeutic antibody. This response may neutralize the therapeutic mAb, interfere with mAb effector function or cause toxicities. We investigated the potential influence of ADA to modify the tumor-binding capability of a tumor-reactive ‘immunocytokine’ (IC), namely, a fusion protein (hu14.18-IL2) consisting of a humanized, tumor-reactive, anti-GD2 mAb genetically linked to interleukin 2. We characterize the role of treatment delivery of IC (intravenous vs intratumoral) on the impact of ADA on therapeutic outcome following IC treatments in an established antimelanoma (MEL) regimen involving radiotherapy (RT) +IC.MethodsC57BL/6 mice were injected with human IgG or the hu14.18-IL2 IC to develop a mouse anti-human antibody (MAHA) response (MAHA+). In vitro assays were performed to assess ADA binding to IC using sera from MAHA+ and MAHA− mice. In vivo experiments assessed the levels of IC bound to tumor in MAHA+ and MAHA− mice, and the influence of IC route of delivery on its ability to bind to B78 (GD2+) MEL tumors.ResultsMAHA is inducible in C57BL/6 mice. In vitro assays show that MAHA is capable of inhibiting the binding of IC to GD2 antigen on B78 cells, resulting in impaired ADCC mediated by IC. When B78-bearing mice are injected intravenously with IC, less IC binds to B78-MEL tumors in MAHA+ mice than in MAHA− mice. In contrast, when IC is injected intratumorally in tumor-bearing mice, the presence of MAHA does not detectibly impact IC binding to the tumor. Combination therapy with RT+IT-IC showed improved tumor regression compared with RT alone in MAHA+ mice. If given intratumorally, IC could be safely readministered in tumor-bearing MAHA+ mice, while intravenous injections of IC in MAHA+ mice caused severe toxicity. Histamine levels were elevated in MAHA+ mice compared with MAHA− mice after reintroduction of IC.ConclusionsIntratumoral injection may be a means of overcoming ADA neutralization of therapeutic activity of tumor-reactive mAbs or ICs and may reduce systemic toxicity, which could have significant translational relevance.

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1820P Taxane-related skin toxicity: Analysis of an in vivo and 3D in vitro model study of the impact of paclitaxel on skin

Annals of Oncology ◽

10.1016/j.annonc.2020.08.1467 ◽

2020 ◽

Vol 31 ◽

pp. S1050

Author(s):

M. Perez-Leal ◽

J.A. Perez Fidalgo ◽

C. Sanz ◽

J. Poveda ◽

J. Milara ◽

...

Keyword(s):

In Vitro Model ◽

Skin Toxicity ◽

Model Study ◽

3D In Vitro Model ◽

Toxicity Analysis ◽

Vitro Model ◽

The Impact

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Rapamycin Reduces Podocyte Apoptosis and is Involved in Autophagy and mTOR/ P70S6K/4EBP1 Signaling

Cellular Physiology and Biochemistry ◽

10.1159/000491905 ◽

2018 ◽

Vol 48 (2) ◽

pp. 765-772 ◽

Cited By ~ 11

Author(s):

Juan Jin ◽

Kang Hu ◽

Meiyu Ye ◽

Diandian Wu ◽

Qiang He

Keyword(s):

Membranous Nephropathy ◽

Annexin V ◽

Cell Counting ◽

Idiopathic Membranous Nephropathy ◽

Therapeutic Effects ◽

Puromycin Aminonucleoside ◽

Group Protein ◽

Vitro Model ◽

The Impact

Background/Aims: The purpose of this study was to investigate the impact of rapamycin (RAP) on autophagy in podocytes and the therapeutic effects of RAP on idiopathic membranous nephropathy (IMN). Methods: We established an in vitro model of IMN by preconditioning mouse podocytes with puromycin aminonucleoside (PAN). A Cell Counting Kit-8 was used to detect the proliferation of each group of podocytes. Podocyte apoptosis was analyzed by flow cytometry via annexin V/propidium iodide dual staining. Subsequently, we observed the number of autophagosomes by transmission electron microscopy. Western blotting was used to detect the levels of LC3, mTOR, p-mTOR, 4EBP1, p-4EBP1, P70S6K, and p-P70S6K in each group. Results: The number of podocytes in the PAN + 100 ng/mL RAP group, PAN + 200 ng/mL RAP group, and PAN + 300 ng/mL RAP group was significantly increased (P < 0.01). The apoptotic rate of podocytes was significantly different between the PAN group and the PAN + RAP group (P < 0.001). There were fewer autophagic corpuscles in the PAN group and more autophagosomes were observed in the PAN + RAP group. LC3 protein expression was down-regulated in the PAN group, while its expression was up-regulated in the PAN + RAP group. In the PAN group, the levels of phosphorylated mTOR, 4EBP1, and P70S6K were increased, while in the PAN + RAP group, protein phosphorylation was reduced. Conclusions: RAP can effectively inhibit the mTOR/P70S6K/4EBP1 signaling pathway, and activate podocyte autophagy, consequently reducing podocyte apoptosis. Therefore, RAP could be used for the treatment of idiopathic membranous nephropathy.

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Spinal cord–fragment interactions following burst fracture: an in vitro model

Journal of Neurosurgery Spine ◽

10.3171/spi.2006.5.3.243 ◽

2006 ◽

Vol 5 (3) ◽

pp. 243-250 ◽

Cited By ~ 19

Author(s):

Richard M. Hall ◽

Robert J. Oakland ◽

Ruth K. Wilcox ◽

David C. Barton

Keyword(s):

Spinal Cord ◽

High Speed ◽

In Vitro Model ◽

Burst Fracture ◽

Posterior Longitudinal Ligament ◽

Bone Fragment ◽

The Status ◽

Vitro Model ◽

The Impact

Object The purpose of the study was to develop an in vitro model of the bone fragment and spinal cord interactions that occur during a burst fracture and further the understanding of how the velocity of the bone fragment and the status of the posterior longitudinal ligament (PLL) affect the deformation of the cord. Methods An in vitro model was developed such that high-speed video and pressure measurements recorded the impact of a simulated bone fragment on sections of explanted bovine spinal cord. The model simulated the PLL and the posterior elements. The status of the PLL had a significant effect on both the maximum occlusion of the spinal cord and the time for occlusion to occur. Raising the fragment velocity led to an overall increase in the spinal cord deformation. Interestingly the dura mater appeared to have little or no effect on the extent of occlusion. Conclusions These findings may indicate the importance of the dura’s interaction with the cerebrospinal fluid in protecting the cord during this type of impact.

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