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BACKGROUND: Metastasis is the leading cause of death in breast cancer patients. For metastasis to occur, tumor cells must invade locally, intravasate, and colonize distant tissues and organs, all steps that require tumor cell migration. The majority of studies on invasion and metastasis rely on human breast cancer cell lines. While it is known that these cells have different properties and abilities for growth and metastasis, the in vitro morphological, proliferative, migratory, and invasive behavior of these cell lines and their correlation to in vivo behavior is poorly understood. Thus, we sought to classify each cell line as poorly or highly metastatic by characterizing tumor growth and metastasis in a murine model of six commonly used human triple-negative breast cancer xenografts, as well as determine which in vitro assays commonly used to study cell motility best predict in vivo metastasis. METHODS: We evaluated the liver and lung metastasis of human TNBC cell lines MDA-MB-231, MDA-MB-468, BT549, Hs578T, BT20, and SUM159 in immunocompromised mice. We characterized each cell line's cell morphology, proliferation, and motility in 2D and 3D to determine the variation in these parameters between cell lines. RESULTS: We identified MDA-MB-231, MDA-MB-468, and BT549 cells as highly tumorigenic and metastatic, Hs578T as poorly tumorigenic and metastatic, BT20 as intermediate tumorigenic with poor metastasis to the lungs but highly metastatic to the livers, and SUM159 as intermediate tumorigenic but poorly metastatic to the lungs and livers. We showed that metrics that characterize cell morphology are the most predictive of tumor growth and metastatic potential to the lungs and liver. Further, we found that no single in vitro motility assay in 2D or 3D significantly correlated with metastasis in vivo. CONCLUSIONS: Our results provide an important resource for the TNBC research community, identifying the metastatic potential of 6 commonly used cell lines. Our findings also support the use of cell morphological analysis to investigate the metastatic potential and emphasize the need for multiple in vitro motility metrics using multiple cell lines to represent the heterogeneity of metastasis in vivo.
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Neoplasias de Mama Triplo Negativas , Humanos , Animais , Camundongos , Neoplasias de Mama Triplo Negativas/patologia , Linhagem Celular Tumoral , Proliferação de Células , Xenoenxertos , Transplante Heterólogo , Movimento CelularRESUMO
BACKGROUND: Individuals who identify as sexual and gender minorities often experience high rates of adverse childhood experiences and encounter discrimination and stigma in their interactions with healthcare providers, leading to low utilization of healthcare services. However, the relationship between adverse childhood experiences, preventive care utilization, and trust in nurses among sexual and gender minority individuals remains unclear. PURPOSE: This study explored the relationship between adverse childhood experiences and preventive care use and assessed the potential interaction effects of trust in nurses between adverse childhood experiences and preventive care use among individuals from sexual and gender minorities. METHODS: A cross-sectional design was used. A sample of 160 self-reported individuals from sexual and gender minorities completed an electronic online survey. Multiple linear regression and moderation analyses were conducted to examine the association between adverse childhood experiences, preventive care utilization, and nurse trust. RESULTS: There was a significant negative relationship between adverse childhood experiences and preventive care utilization. There was also a significant positive relationship between trust and preventive care utilization. The results also indicated that trust in nurses moderated the relationship between adverse childhood experiences and preventive care utilization among sexual and gender minority individuals. DISCUSSION: A significant relationship was found between a high prevalence of adverse childhood experiences and low healthcare service utilization. Strengthening the trust relationship between nurses and sexual and gender minority individuals could serve as a potential intervention point, leading to improved health outcomes for this vulnerable population. Hence, enhancing trust in nurses could be a key factor in increasing healthcare service utilization and overall health outcomes.
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Experiências Adversas da Infância , Minorias Sexuais e de Gênero , Humanos , Estudos Transversais , Comportamento Sexual , Confiança , Masculino , FemininoRESUMO
Acute myeloid leukemia (AML) is a major unmet medical need. Most patients have poor long-term survival, and treatment has not significantly changed in 40 years. Recently, bispecific antibodies that redirect the cytotoxic activity of effector T cells by binding to CD3, the signaling component of the T-cell receptor, and a tumor target have shown clinical activity. Notably, blinatumomab is approved to treat relapsed/refractory acute lymphoid leukemia. Here we describe the design, discovery, pharmacologic activity, pharmacokinetics, and safety of a CD3 T cell-dependent bispecific (TDB) full-length human IgG1 therapeutic antibody targeting CLL-1 that could potentially be used in humans to treat AML. CLL-1 is prevalent in AML and, unlike other targets such as CD33 and CD123, is not expressed on hematopoietic stem cells providing potential hematopoietic recovery. We selected a high-affinity monkey cross-reactive anti-CLL-1 arm and tested several anti-CD3 arms that varied in affinity, and determined that the high-affinity CD3 arms were up to 100-fold more potent in vitro. However, in mouse models, the efficacy differences were less pronounced, probably because of prolonged exposure to TDB found with lower-affinity CD3 TDBs. In monkeys, assessment of safety and target cell depletion by the high- and low-affinity TDBs revealed that only the low-affinity CD3/CLL1 TDB was well tolerated and able to deplete target cells. Our data suggest that an appropriately engineered CLL-1 TDB could be effective in the treatment of AML.
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Anticorpos Biespecíficos/uso terapêutico , Antineoplásicos/uso terapêutico , Lectinas Tipo C/imunologia , Leucemia Mieloide Aguda/tratamento farmacológico , Lectina 3 Semelhante a Ig de Ligação ao Ácido Siálico/imunologia , Animais , Anticorpos Biespecíficos/efeitos adversos , Anticorpos Biespecíficos/imunologia , Anticorpos Biespecíficos/farmacocinética , Antineoplásicos/efeitos adversos , Antineoplásicos/imunologia , Antineoplásicos/farmacocinética , Linhagem Celular Tumoral , Humanos , Leucemia Mieloide Aguda/imunologia , Leucemia Mieloide Aguda/patologia , Macaca fascicularis , Camundongos Endogâmicos C57BL , Camundongos TransgênicosRESUMO
BACKGROUND: Symptom index (SI) and symptom association probability (SAP) are popular methods used to measure symptom association in patients with gastroesophageal reflux disease (GERD). AIM: To investigate whether these 2 methods yield similar results in analysis of both typical and atypical GERD symptoms. MATERIALS AND METHODS: Combined impedance-pH reflux studies of 1471 patients tested for possible GERD symptoms from January 2010 to May 2015 were reviewed. SI and SAP were analyzed for typical and atypical GERD symptoms including heartburn, regurgitation, indigestion, chest pain, cough, and throat clearing (TC). Patients who reported <3 symptom events during the 24-hour monitoring period were excluded. ON and OFF proton pump inhibitor (PPI) groups were reviewed. Kappa coefficient (κ) rather than simple percentage was used to measure the agreement rate. Simple percentage agreement is a less reliable method compared with κ. RESULTS: On PPI therapy, there was a good κ between SI and SAP for regurgitation (0.68) and indigestion (0.64), moderate for heartburn (0.48) and chest pain (0.51), and poor for cough (0.33) and TC (0.29). There was a lower κ OFF PPI therapy for heartburn (0.36), regurgitation (0.44), and indigestion (0.50). But there was no difference in κ for chest pain (0.61), cough (0.29), and TC (0.33). CONCLUSIONS: SI and SAP showed better agreement for patients with typical GERD symptoms and even better when tested ON PPI. A better symptom association method is needed for patients with atypical GERD symptoms.
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Refluxo Gastroesofágico/diagnóstico , Índice de Gravidade de Doença , Impedância Elétrica , Monitoramento do pH Esofágico , Feminino , Refluxo Gastroesofágico/tratamento farmacológico , Refluxo Gastroesofágico/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Inibidores da Bomba de Prótons/uso terapêuticoRESUMO
PURPOSE: THIOMAB™ drug conjugates (TDCs) with engineered cysteine residues allow site-specific drug conjugation and defined Drug-to-Antibody Ratios (DAR). In order to help elucidate the impact of drug-loading, conjugation site, and subsequent deconjugation on pharmacokinetics and efficacy, we have developed an integrated mathematical model to mechanistically characterize pharmacokinetic behavior and preclinical efficacy of MMAE conjugated TDCs with different DARs. General applicability of the model structure was evaluated with two different TDCs. METHOD: Pharmacokinetics studies were conducted for unconjugated antibody and purified TDCs with DAR-1, 2 and 4 for trastuzumab TDC and Anti-STEAP1 TDC in mice. Total antibody concentrations and individual DAR fractions were measured. Efficacy studies were performed in tumor-bearing mice. RESULTS: An integrated model consisting of distinct DAR species (DAR0-4), each described by a two-compartment model was able to capture the experimental data well. Time series measurements of each Individual DAR species allowed for the incorporation of site-specific drug loss through deconjugation and the results suggest a higher deconjugation rate from heavy chain site HC-A114C than the light chain site LC-V205C. Total antibody concentrations showed multi-exponential decline, with a higher clearance associated with higher DAR species. The experimentally observed effects of TDC on tumor growth kinetics were successfully described by linking pharmacokinetic profiles to DAR-dependent killing of tumor cells. CONCLUSION: Results from the integrated model evaluated with two different TDCs highlight the impact of DAR and site of conjugation on pharmacokinetics and efficacy. The model can be used to guide future drug optimization and in-vivo studies.
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Anticorpos Monoclonais/farmacocinética , Antineoplásicos/farmacocinética , Modelos Biológicos , Compostos de Sulfidrila/farmacocinética , Trastuzumab/metabolismo , Administração Intravenosa , Animais , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais/química , Antígenos de Neoplasias/imunologia , Antineoplásicos/administração & dosagem , Antineoplásicos/química , Disponibilidade Biológica , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Cisteína , Feminino , Masculino , Taxa de Depuração Metabólica , Camundongos Nus , Camundongos SCID , Transplante de Neoplasias , Compostos de Sulfidrila/administração & dosagem , Compostos de Sulfidrila/química , Trastuzumab/administração & dosagem , Trastuzumab/químicaRESUMO
The fear of COVID-19 significantly impacting the health of people globally. This study translated newly developed measurement tool New Fear of the Coronavirus Questionnaire (New_FCQ) into Chinese language and evaluated the psychometric properties of the Chinese version of New_FCQ among Chinese population. A total of 522 participants were included in the study. Internal consistency, construct validity, criterion validity, and concurrent validity of the Chinese version of New_FCQ were assessed in this study. The Chinese version of New_FCQ had excellent internal consistency (αâ =â 0.97) and exploratory factor analysis demonstrated one-dimensional structure of the Chinese version of New_FCQ. The preliminary criterion validity revealed statistically significant differences in the fear of COVID-19 scores based on age and education level (Pâ =â .002 and Pâ =â .03, respectively). The good concurrent validity also established with the Chinese version Fear of COVID-19 Scale(Pâ <â .001). Psychometric proportions of the Chinese version of New_FCQ were established, which exhibited sufficient validity and reliability among Chinese population.
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COVID-19 , Coronavirus , Humanos , Psicometria/métodos , Reprodutibilidade dos Testes , Medo , Inquéritos e Questionários , IdiomaRESUMO
Obesity is associated with increased incidence and metastasis of triple-negative breast cancer (TNBC), an aggressive breast cancer subtype. The extracellular matrix (ECM) is a major component of the tumor microenvironment that drives metastasis. To characterize the temporal effects of age and high-fat diet-driven weight gain on the ECM, we injected allograft tumor cells at 4-week intervals into mammary fat pads of mice fed a control or high-fat diet (HFD), assessing tumor growth and metastasis and evaluating the ECM composition of the mammary fat pads, lungs, and livers. Tumor growth was increased in obese mice after 12 weeks on the HFD. Liver metastasis increased in obese mice only at 4 weeks, and elevated body weight correlated with increased metastasis to the lungs but not the liver. Whole decellularized ECM coupled with proteomics indicated that early stages of obesity were sufficient to induce changes in the ECM composition. Obesity led to increased abundance of the pro-invasive ECM proteins collagen IV and collagen VI in the mammary glands and enhanced the invasive capacity of cancer cells. Cells of stromal vascular fraction and adipose stem and progenitor cells were primarily responsible for secreting collagen IV and VI, not adipocytes. Longer exposure to HFD increased the invasive potential of ECM isolated from lung and liver, with significant changes in ECM composition found in the liver with short-term HFD exposure. Together, this data suggests that changes in the breast, lung, and liver ECM underlie some of the effects of obesity on TNBC incidence and metastasis.
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Antibody-drug conjugates (ADCs) are target-specific anticancer agents consisting of cytotoxic drugs covalently linked to a monoclonal antibody. The number of ADCs in the clinic is growing, and therefore thorough characterization of the quantitative assays used to measure ADC concentrations in support of pharmacokinetic, efficacy, and safety studies is of increasing importance. Cytotoxic drugs such as the tubulin polymerization inhibiting auristatin, monomethyl auristatin E, have been conjugated to antibodies via cleavable linkers (MC-vc-PAB) through internal cysteines. This results in a heterogeneous mixture of antibody species with drug-to-antibody ratios (DAR) ranging from 0 to 8. In order to characterize the assays used to quantitate total MC-vc-PAB-MMAE ADCs (conjugated and unconjugated antibody), we used purified fractions with defined DARs from 6 therapeutic antibodies to evaluate different assay formats and reagents. Our investigations revealed that for quantitation of total antibody, including all unconjugated and conjugated antibody species, sandwich ELISA formats did not always allow for recovery of all purified DAR fractions (DAR 0-8) to within ±20% of the expected values at the reagent concentrations tested. In evaluating alternative approaches, we found that the recovery of DAR fractions with semihomogeneous assay (SHA) formats, in which sample, capture, and detection reagents are preincubated in solution, were less affected by the antibody's MMAE drug load as compared to traditional stepwise sandwich ELISAs. Thus, choosing the optimal assay format and reagents for total antibody assays is valuable for developing accurate quantitative assays.
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Antineoplásicos/farmacocinética , Imunotoxinas/farmacocinética , Oligopeptídeos/farmacocinética , Moduladores de Tubulina/farmacocinética , Animais , Antineoplásicos/química , Ensaio de Imunoadsorção Enzimática , Imunotoxinas/química , Camundongos , Camundongos SCID , Oligopeptídeos/química , Moduladores de Tubulina/químicaRESUMO
INTRODUCTION AND IMPORTANCE: Necrotizing fasciitis is an aggressive skin and soft tissue infection that is a surgical emergency, and Haemophilus influenzae (H. flu) is a rare cause. We present a case of H. flu co-infection causing necrotizing fasciitis in the setting of COVID-19 pneumonia. CASE PRESENTATION: A 56-year-old male presented with 2 weeks of upper respiratory symptoms. He was unvaccinated against COVID-19 and tested positive for COVID-19 five days prior. He developed respiratory failure requiring intubation, and was treated with dexamethasone, remdesivir, and tocilizumab for COVID-19 pneumonia. On hospital day 2, he was hypotensive with new rapidly evolving erythematous lesions with crepitus of his lower extremities suspicious for necrotizing fasciitis. He underwent wide excision and debridement with significant hemodynamic improvements. H. flu co-infection was identified from blood cultures. Aberrant cells with 94 % lymphocytes were noted and suggested chronic lymphocytic leukemia (CLL) that was not previously known. He developed progressive lesions globally, concerning for purpura fulminans with clinical disseminated intravascular coagulation and neurological decline ultimately leading to withdrawal of care. CLINICAL DISCUSSION: COVID-19 infection is often associated with concomitant opportunistic infections. Our patient was also immunocompromised by CLL, diabetes, chronic steroids, and initial appropriate COVID-19 treatments. Despite appropriate treatments, he could not overcome his medical comorbidities and multiple infections. CONCLUSION: Necrotizing fasciitis caused by H. flu is rare, and we present the first case as a co-infection in the setting of COVID-19 pneumonia. Due to the patient's immunocompromised state with underlying CLL, this proved to be fatal.
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PURPOSE: Our study investigates whether preoperative anterior chamber depth (ACD) measured by Scheimpflug tomography could serve as a potential predictor of graft failure in eyes undergoing Descemet stripping endothelial keratoplasty (DSEK). METHODS: A retrospective review was conducted on patients who underwent primary or repeat DSEK between January 2020 and August 2021 at Bascom Palmer Eye Institute. Charts from 378 primary and 192 repeat DSEK patients were reviewed and ultimately 47 primary and 21 repeat DSEK patients met criteria for inclusion. Data collection included demographics, preoperative ACD, best-corrected visual acuity, and length of follow-up. RESULTS: Demographics were similar between groups, and there was no significant difference in the average best-corrected visual acuity between the single and repeat DSEK groups preoperatively. Baseline preoperative ACD was greatest in the single DSEK group (3.51 ± 0.90 mm) when compared to baseline preoperative ACD in the repeat DSEK group (3.01 ± 0.67 mm, P = 0.003). The preoperative mean ACD was smallest in the repeat DSEK group before the second DSEK (2.94 ± 0.48 mm, P = 0.001). Preoperative baseline ACD was the only variable to affect graft survival time significantly (P = 0.012). The incidence of glaucoma diagnosis was similar in both groups (42.5% vs. 42.8%, P = 0.471). The diagnosis of glaucoma and presence of incisional glaucoma surgery did not affect the graft survival time (P = 0.129, P = 0.559) or need for repeat DSEK. CONCLUSIONS: Smaller baseline preoperative Scheimpflug ACD measurement may be a possible predictor of the need for repeat DSEK. Our study found that Scheimpflug ACD decreases with subsequent DSEK failure.
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BACKGROUND: Residents are often viewed as contributors to Emergency Department (ED) prolongation of length of stay (LOS). To understand this proposition, we performed a study to identify ED patient care intervals and how each contributed to LOS. METHODS: We performed a retrospective review of prospectively gathered data on 145 ED surgery consults. Residents prospectively documented patient names, page times, and time of plan. Key ED patient care intervals were then retrospectively extracted from the patient's chart. A time analysis was then performed. RESULTS: Average arrival to disposition time was 305 min, and residents averaged 47 min to see and staff consults. The longest intervals were arrival to imaging (75 min) and imaging time (73 min). Average disposition to discharge time was 170 min (36% of LOS). CONCLUSIONS: Surgery residents see and staff consults within the norms for care established by the hospital. Imaging time is a bottleneck hindering disposition. Access block also significantly increases ED LOS.
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Internato e Residência , Serviço Hospitalar de Emergência , Humanos , Tempo de Internação , Encaminhamento e Consulta , Estudos RetrospectivosRESUMO
Antibody-drug conjugates (ADCs) are designed to combine the exquisite specificity of antibodies to target tumor antigens with the cytotoxic potency of chemotherapeutic drugs. In addition to the general chemical stability of the linker, a thorough understanding of the relationship between ADC composition and biological disposition is necessary to ensure that the therapeutic window is not compromised by altered pharmacokinetics (PK), tissue distribution, and/or potential organ toxicity. The six-transmembrane epithelial antigen of prostate 1 (STEAP1) is being pursued as a tumor antigen target. To assess the role of ADC composition in PK, we evaluated plasma and tissue PK profiles in rats, following a single dose, of a humanized anti-STEAP1 IgG1 antibody, a thio-anti-STEAP1 (ThioMab) variant, and two corresponding thioether-linked monomethylauristatin E (MMAE) drug conjugates modified through interchain disulfide cysteine residues (ADC) and engineered cysteines (TDC), respectively. Plasma PK of total antibody measured by enzyme-linked immunosorbent assay (ELISA) revealed â¼45% faster clearance for the ADC relative to the parent antibody, but no apparent difference in clearance between the TDC and unconjugated parent ThioMab. Total antibody clearances of the two unconjugated antibodies were similar, suggesting minimal effects on PK from cysteine mutation. An ELISA specific for MMAE-conjugated antibody indicated that the ADC cleared more rapidly than the TDC, but total antibody ELISA showed comparable clearance for the two drug conjugates. Furthermore, consistent with relative drug load, the ADC had a greater magnitude of drug deconjugation than the TDC in terms of free plasma MMAE levels. Antibody conjugation had a noticeable, albeit minor, impact on tissue distribution with a general trend toward increased hepatic uptake and reduced levels in other highly vascularized organs. Liver uptakes of ADC and TDC at 5 days postinjection were 2-fold and 1.3-fold higher, respectively, relative to the unmodified antibodies. Taken together, these results indicate that the degree of overall structural modification in anti-STEAP1-MMAE conjugates has a corresponding level of impact on both PK and tissue distribution.
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Anticorpos Monoclonais/química , Anticorpos Monoclonais/farmacocinética , Antígenos de Neoplasias/imunologia , Imunoconjugados/química , Imunoconjugados/farmacocinética , Oligopeptídeos/química , Oligopeptídeos/farmacocinética , Oxirredutases/imunologia , Animais , Anticorpos Monoclonais/sangue , Dissulfetos/química , Humanos , Imunoconjugados/sangue , Masculino , Modelos Moleculares , Oligopeptídeos/sangue , Ratos , Ratos Sprague-Dawley , Distribuição TecidualRESUMO
A theoretical safety concern proposed in the influenza literature is that therapeutic antiviral antibodies could have the potential for antibody-dependent enhancement (ADE) of infection and disease. ADE may occur when virus-specific antibodies at subtherapeutic, nonneutralizing concentrations facilitate virus uptake and, in some cases, enhance replication, which can lead to an exacerbation of virus-mediated disease. Alternatively, ADE may occur due to antibody-dependent complement activation exacerbating virus-mediated disease in the absence of increased replication. As a result of this theoretical safety concern, safety assessment of anti-influenza antibodies may include an in vivo evaluation of ADE of infection and/or disease. These studies were conducted to investigate the potential of MHAB5553A, a broadly specific, neutralizing therapeutic anti-influenza B antibody, to elicit ADE of infection and disease in mouse models of influenza B infection. In parallel studies, female DBA/2J mice were infected with either influenza B/Victoria/504/2000 or influenza B/Brisbane/60/2008 representing distinct lineages. Assessment of ADE was based on an integration of results from multiple endpoints, including infectious lung viral titers and genomes, body weight, mortality, lung weight, and histopathology. In these studies, the high dose of 15 mg/kg MHAB5553A resulted in substantial attenuation of influenza pneumonia, with more modest effects at 1.5 mg/kg; whereas MHAB5553A treatment at 0.15 or 0.015 mg/kg was generally comparable to vehicle-treated controls. Our results demonstrate that MHAB5553A across a broad range of doses did not enhance primary influenza B infection or disease in this model, and represent a nonclinical de-risking of the ADE potential with this antibody.
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Anticorpos Monoclonais Humanizados/efeitos adversos , Anticorpos Facilitadores , Vírus da Influenza B/imunologia , Infecções por Orthomyxoviridae/tratamento farmacológico , Animais , Peso Corporal , Relação Dose-Resposta a Droga , Feminino , Genoma Viral , Pulmão/patologia , Pulmão/virologia , Camundongos , Camundongos Endogâmicos DBA , Infecções por Orthomyxoviridae/patologia , Infecções por Orthomyxoviridae/virologiaRESUMO
Prediction of human pharmacokinetics (PK) based on preclinical information for antibody-drug conjugates (ADCs) provide important insight into first-in-human (FIH) study design. This retrospective analysis was conducted to identify an appropriate scaling method to predict human PK for ADCs from animal PK data in the linear range. Different methods for projecting human clearance (CL) from animal PK data for 11 ADCs exhibiting linear PK over the tested dose ranges were examined: multiple species allometric scaling (CL vs. body weight), allometric scaling with correction factors, allometric scaling based on rule of exponent, and scaling from only cynomolgus monkey PK data. Two analytes of interest for ADCs, namely total antibody and conjugate (measured as conjugated drug or conjugated antibody), were assessed. Percentage prediction errors (PEs) and residual sum of squares (RSS) were compared across methods. Human CL was best estimated using cynomolgus monkey PK data alone and an allometric scaling exponent of 1.0 for CL. This was consistently observed for both conjugate and total antibody analytes. Other scaling methods either underestimated or overestimated human CL, or produced larger average absolute PEs and RSS. Human concentration-time profiles were also reasonably predicted from the cynomolgus monkey data using species-invariant time method with a fixed exponent of 1.0 for CL and 1.0 for volume of distribution. In conclusion, results from this retrospective analysis of 11 ADCs indicate that allometric scaling of CL with an exponent of 1.0 using cynomolgus monkey PK data alone can successfully project human PK profiles of an ADC within linear range.
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Imunoconjugados/farmacocinética , Animais , Anticorpos Monoclonais/farmacocinética , Humanos , Macaca fascicularis , Taxa de Depuração MetabólicaRESUMO
TENB2, a transmembrane proteoglycan protein, is a promising target for antibody drug conjugate (ADC) therapy due to overexpression in human prostate tumors and rapid internalization. We previously characterized how predosing with parental anti-TENB2 monoclonal antibody (mAb) at 1 mg/kg in a patient-derived LuCap77 explant model with high (3+) TENB2 expression could (i) block target-mediated intestinal uptake of tracer (& 0.1 mg/kg) levels of radiolabeled anti-TENB2-monomethyl auristatin E ADC while preserving tumor uptake, and (ii) maintain efficacy relative to ADC alone. Here, we systematically revisit this strategy to evaluate the effects of predosing on tumor uptake and efficacy in LuCap96.1, a low TENB2-expressing (1+) patient-derived model that is more responsive to ADC therapy than LuCap77. Importantly, rather than using tracer (& 0.1 mg/kg) levels, radiolabeled ADC tumor uptake was assessed at 1 mg/kg - one of the doses evaluated in the tumor growth inhibition study - in an effort to bridge tissue distribution (PK) with efficacy (PD). Predosing with mAb up to 1 mg/kg had no effect on efficacy. These findings warrant further investigations to determine whether predosing prior to ADC therapy might improve therapeutic index by preventing ADC disposition and possible toxicological liabilities in antigen-expressing healthy tissues.
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Antibody drug conjugates (ADC), in which small molecule cytotoxic agents are non-specifically linked to antibodies, can enable targeted delivery of chemotherapeutics to tumor cells. ADCs are often produced and administered as a mixture of conjugated antibodies with different drug to antibody ratios (DAR) resulting in complex and heterogeneous disposition kinetics. We developed a mechanism-based platform model that can describe and predict the complex pharmacokinetic (PK) behavior of ADCs with protease-cleavable valine-citrulline (VC) linker linked to Monomethylmonomethyl auristatin F/E by incorporating known mechanisms of ADC disposition. The model includes explicit representation of all DAR species; DAR-dependent sequential deconjugation of the drug, resulting in the conversion of higher DAR to lower DAR species; and DAR-dependent antibody/ADC clearance. PK profiles of multiple analytes (total antibody, drug-conjugated antibody, and/or antibody-conjugated drug) for different ADC molecules and targets in rodents and cynomolgus monkeys were used for model development. The integrated cross-species model was successful in capturing the multi-analyte PK profiles after administration of purified ADCs with defined DAR species and ADCs with mixtures of DAR. Human PK predictions for DSTP3086S (anti-STEAP1-vc-MMAE) with the platform model agreed well with PK (total antibody and antibody-conjugated drug concentrations) measurements in the dose-ranging phase I clinical study. The integrated model is applicable to various other ADCs with different formats, conjugated drugs, and linkers, and provides a valuable tool for the exploration of mechanisms governing disposition of ADCs and enables translational predictions.
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Anticorpos Monoclonais/farmacocinética , Antineoplásicos/farmacocinética , Imunoconjugados/farmacocinética , Modelos Biológicos , Oligopeptídeos/farmacocinética , Animais , Simulação por Computador , Humanos , Oligopeptídeos/química , Pesquisa Translacional BiomédicaRESUMO
PURPOSE: Antibody-drug conjugates (ADC) selectively deliver a cytotoxic drug to cells expressing an accessible antigenic target. Here, we have appended monomethyl auristatin E (MMAE) to an antibody recognizing the SLC34A2 gene product NaPi2b, the type II sodium-phosphate cotransporter, which is highly expressed on tumor surfaces of the lung, ovary, and thyroid as well as on normal lung pneumocytes. This study evaluated its efficacy and safety in preclinical studies. EXPERIMENTAL DESIGN: The efficacy of anti-NaPi2b ADC was evaluated in mouse ovarian and non-small cell lung cancer (NSCLC) tumor xenograft models, and its toxicity was assessed in rats and cynomolgus monkeys. RESULTS: We show here that an anti-NaPi2b ADC is effective in mouse ovarian and NSCLC tumor xenograft models and well-tolerated in rats and cynomolgus monkeys at levels in excess of therapeutic doses. Despite high levels of expression in normal lung of non-human primate, the cross-reactive ADC exhibited an acceptable safety profile with a dose-limiting toxicity unrelated to normal tissue target expression. The nonproliferative nature of normal pneumocytes, together with the antiproliferative mechanism of MMAE, likely mitigates the potential liability of this normal tissue expression. CONCLUSIONS: Overall, our preclinical results suggest that the ADC targeting NaPi2b provides an effective new therapy for the treatment of NSCLC and ovarian cancer and is currently undergoing clinical developments.
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Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Oligopeptídeos/administração & dosagem , Neoplasias Ovarianas/tratamento farmacológico , Proteínas Cotransportadoras de Sódio-Fosfato Tipo IIb/imunologia , Animais , Anticorpos Monoclonais/administração & dosagem , Carcinoma Pulmonar de Células não Pequenas/imunologia , Carcinoma Pulmonar de Células não Pequenas/patologia , Feminino , Humanos , Imunoconjugados/administração & dosagem , Macaca fascicularis , Masculino , Camundongos , Oligopeptídeos/imunologia , Neoplasias Ovarianas/imunologia , Neoplasias Ovarianas/patologia , Ratos , Proteínas Cotransportadoras de Sódio-Fosfato Tipo IIb/antagonistas & inibidores , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Miniaturization of immunoassays has numerous potential advantages over traditional ELISAs. Here we present a novel approach using patterned planar plates (PPPs). These 'wall-less' plates consist of a 16 × 24 array of 2 mm diameter hydrophilic regions surrounded by a hydrophobic polytetrafluoroethylene (PTFE) coating. Assays are performed by adding 2 µL droplets to the hydrophilic areas. These droplets are overlaid with an immiscible mixture of perfluorocarbon liquid (PFCL) that essentially eliminates evaporation. During wash steps, a thin film of PFCL covers the hydrophobic coating and prevents its wetting by wash buffer; as a result, the hydrophilic wells remain intact and inter-well cross-contamination is prevented. We compared the performance of three immunoassays using PPPs versus traditional 384-well ELISA plates. These included assays for soluble FcRH5 in human serum, SDF-1 in mouse serum, and human IgG in mouse plasma. The results show that the PPP assays were closely comparable to the ELISAs in terms of sensitivity, linearity of dilution, and sample quantitation. Moreover, the PPP assays were rapid to perform, easily adapted from ELISA protocols, and used 10- to 50-fold less sample and reagent volume as compared to 384- or 96-well plate ELISAs. As an additional advantage, PPPs conform to established microplate dimensional standards making them compatible with pre-existing equipment and workflows. PPPs therefore represent an attractive and broadly applicable approach to flexible miniaturization of plate-based immunochemical assays.
Assuntos
Imunoensaio/instrumentação , Análise em Microsséries/instrumentação , Animais , Anticorpos Monoclonais/sangue , Anticorpos Monoclonais/imunologia , Quimiocina CXCL12/análise , Quimiocina CXCL12/imunologia , Humanos , Interações Hidrofóbicas e Hidrofílicas , Imunoglobulina G/imunologia , Camundongos , Politetrafluoretileno/química , Receptores de Superfície Celular/análise , Receptores de Superfície Celular/imunologia , Receptores FcRESUMO
BACKGROUND AND PURPOSE: The success of antibody-drug conjugates (ADCs) depends on the therapeutic window rendered by the differential expression between normal and pathological tissues. The ability to identify and visualize target expression in normal tissues could reveal causes for target-mediated clearance observed in pharmacokinetic characterization. TENB2 is a prostate cancer target associated with the progression of poorly differentiated and androgen-independent tumour types, and ADCs specific for TENB2 are candidate therapeutics. The objective of this study was to locate antigen expression of TENB2 in normal tissues, thereby elucidating the underlying causes of target-mediated clearance. EXPERIMENTAL APPROACH: A series of pharmacokinetics, tissue distribution and mass balance studies were conducted in mice using a radiolabelled anti-TENB2 ADC. These data were complemented by non-invasive single photon emission computed tomography - X-ray computed tomography imaging and immunohistochemistry. KEY RESULTS: The intestines were identified as a saturable and specific antigen sink that contributes, at least in part, to the rapid target-mediated clearance of the anti-TENB2 antibody and its drug conjugate in rodents. As a proof of concept, we also demonstrated the selective disposition of the ADC in a tumoural environment in vivo using the LuCaP 77 transplant mouse model. High tumour uptake was observed despite the presence of the antigen sink, and antigen specificity was confirmed by antigen blockade. CONCLUSIONS AND IMPLICATIONS: Our findings provide the anatomical location and biological interpretation of target-mediated clearance of anti-TENB2 antibodies and corresponding drug conjugates. Further investigations may be beneficial in addressing the relative contributions to ADC disposition from antigen expression in both normal and pathological tissues.