Measuring phospho-MET by multiplex immunofluorescence to aid in selection of patients with MET activation in tumors.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. 3131-3131
Author(s):  
Tony Navas ◽  
Apurva K. Srivastava ◽  
Jeevan P Govindharajulu ◽  
Yvonne A. Evrard ◽  
Susanne Borgel ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Patient Selection ◽  
Tumor Tissue ◽  
Tissue Microarrays ◽  
Immunofluorescence Assay ◽  
Preclinical Trials ◽  
Xenograft Models ◽  
Patient Selection Criteria ◽  
Development And Validation ◽  
Pdx Models

3131 Background: Currently, patient selection criteria for clinical testing of MET inhibitors are limited. Robust studies selecting patients based on MET protein expression, MET gene amplification, or mutations have not met their efficacy goals. Development of microscopy-based assays to quantify levels of phospho-MET (pMET) in tumors has been hampered by poor antibody specificity. Here, we present the development and validation of a robust, highly specific multiplex immunofluorescence assay (IFA) that measures pY1235-MET and total MET in tumor tissue. Methods: This assay utilizes antibodies to pY1235-MET (NCI-23111), total MET (D1C2), and plasma membrane (PM) marker Na+/K+-ATPase, each conjugated to a different Alexa Fluor dye. We used tumor tissue from crizotinib-treated SNU5 xenograft models to demonstrate pY1235-MET assay fitness-for-purpose and cross-platform assay concordance with our validated pMET ELISA. In addition, this IFA was validated by phospho-peptide competition using custom tissue microarrays (TMA) derived from patients with colorectal carcinoma (CRC). Finally, we developed quantitative algorithms to assess pY1235 MET levels in the plasma membrane and nucleus using PM and DAPI masks, respectively. Patient-derived xenograft models (PDX) were obtained from NCI’s Patient-Derived Models Repository (www.pdmr.cancer.gov). Results: The prevalence of high pY1235-MET expression in CRC patient specimens was greater than expected; of the 64 TMA cores evaluated, 29 (45%) and 19 (29%) had high pY1235-MET and total MET levels, respectively, as defined by mean marker area of ≥ 30 μm2/cell. To address the potential utility of pY1235-MET as a diagnostic biomarker, we examined 15 CRC PDX models by pMET ELISA and IFA. Two CRC tumor models were positive for pY1235-MET expression in both assays. The pY1235-MET IFA results and gene expression data were used to select PDX models for ongoing preclinical trials of potent MET inhibitors. Conclusions: This novel pY1235-MET IFA will enable clinicians to address the utility of activated MET as a biomarker for patient selection and/or prediction of response in clinical trials of MET inhibitors. Funded by NCI Contract No. HHSN261200800001E.

scholarly journals Patient-Derived Tumor Xenograft Models: Toward the Establishment of Precision Cancer Medicine

2020 ◽  
Vol 10 (3) ◽  
pp. 64
Author(s):  
Taichiro Goto
Keyword(s):  
Tumor Tissue ◽  
Tumor Xenograft ◽  
Preclinical Models ◽  
Immunodeficient Mice ◽  
Personalized Cancer Medicine ◽  
Novel Treatment ◽  
Xenograft Models ◽  
Cancer Medicine ◽  
Personalized Cancer ◽  
Pdx Models

Patient-derived xenografts (PDXs) describe models involving the implantation of patient-derived tumor tissue into immunodeficient mice. Compared with conventional preclinical models involving the implantation of cancer cell lines into mice, PDXs can be characterized by the preservation of tumor heterogeneity, and the tumor microenvironment (including stroma/vasculature) more closely resembles that in patients. Consequently, the use of PDX models has improved the predictability of clinical therapeutic responses to 80% or greater, compared with approximately 5% for existing models. In the future, molecular biological analyses, omics analyses, and other experiments will be conducted using recently prepared PDX models under the strong expectation that the analysis of cancer pathophysiology, stem cells, and novel treatment targets and biomarkers will be improved, thereby promoting drug development. This review outlines the methods for preparing PDX models, advances in cancer research using PDX mice, and perspectives for the establishment of precision cancer medicine within the framework of personalized cancer medicine.

PATIENT SELECTION CRITERIA IN THE SURGICAL TREATMENT OF VENO-OCCLUSIVE DYSFUNCTION

1999 ◽  
Vol 161 (4) ◽  
pp. 1145-1147 ◽  
Author(s):  
F. SASSO ◽  
G. GULINO ◽  
J. WEIR ◽  
A.M. VIGGIANO ◽  
E. ALCINI
Keyword(s):  
Surgical Treatment ◽  
Patient Selection ◽  
Selection Criteria ◽  
Patient Selection Criteria

scholarly journals Suggested Patient Selection Criteria for Initial Clinical Trials of Pig Kidney Xenotransplantation in the USA

2021 ◽  
Vol Publish Ahead of Print ◽  
Author(s):  
Abhijit Jagdale ◽  
Vineeta Kumar ◽  
Douglas J. Anderson ◽  
Jayme E. Locke ◽  
Michael J. Hanaway ◽  
...  
Keyword(s):  
Clinical Trials ◽  
Patient Selection ◽  
Selection Criteria ◽  
Pig Kidney ◽  
The Usa ◽  
Patient Selection Criteria

scholarly journals High Prevalence of 5T4/Trophoblast Glycoprotein in Soft Tissue Sarcomas

Cancers ◽  
2021 ◽  
Vol 13 (19) ◽  
pp. 4841
Author(s):  
Patrick Groothuis ◽  
Nicola Penel ◽  
Antoine Italiano ◽  
Nuria Kotecki ◽  
Fred Dijcks ◽  
...  
Keyword(s):  
Soft Tissue ◽  
Tumor Tissue ◽  
Staining Method ◽  
Soft Tissue Sarcomas ◽  
Tissue Microarrays ◽  
Histological Subtypes ◽  
Scoring Method ◽  
Drug Conjugates ◽  
High Prevalence ◽  
Quantitative Scoring

The expression of 5T4/trophoblast glycoprotein was evaluated in several histological subtypes of soft tissue sarcoma (STS) to determine whether the prevalence and level of expression of this membrane-associated glycoprotein is sufficient for use in targeted therapies. Tumor tissue microarrays containing cores from different histological subtypes of STS were stained using a standardized immunohistochemical staining method to detect 5T4; the level of staining was assessed using a semi-quantitative scoring method. No 5T4 staining was seen in the angiosarcomas and liposarcomas investigated in this study. 5T4 staining in the other STS subtypes was seen in more than 50% of cases, warranting further investigation into whether this antigen could evoke an anti-tumor immune response or can be used as target for the delivery of more potent toxins through antibody drug conjugates.

EXTH-09. TUMOR PHARMACOKINETICS, PHARMACODYNAMICS AND RADIATION SENSITIZATION IN PATIENT-DERIVED XENOGRAFT MODELS OF GLIOBLASTOMA TREATED WITH THE AURORA KINASE A INHIBITOR LY3295668

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi165-vi165
Author(s):  
Shwetal Mehta ◽  
Artak Tovmasyan ◽  
An-Chi Tien ◽  
Michael Holter ◽  
Barbara Hopkins ◽  
...  
Keyword(s):  
Tumor Tissue ◽  
Synergistic Effects ◽  
Equilibrium Dialysis ◽  
Aurora Kinase ◽  
Intracranial Tumors ◽  
Aurora Kinase A ◽  
Patient Derived Xenograft ◽  
Radiation Sensitization ◽  
Pdx Models ◽  
Kinase A

Abstract Cell-cycle deregulation is at the crux of all malignancies, including glioblastoma (GBM). Aurora Kinase A (AURKA) plays a central role in G2/M transition and faithful chromosome segregation. In this study, we evaluated the pharmacokinetics, pharmacodynamics, and radiation sensitization properties of LY3295668, a highly specific AURKA inhibitor, in orthotopic patient-derived xenograft (PDX) models of GBM. Mice with intracranial tumors were randomized to 50 mg/kg LY3295668 PO BID x 4 days vs. placebo. LY3295668 levels in plasma and contrast-enhancing tumor tissue were measured by liquid chromatography tandem mass spectrometry (LC-MS/MS). Unbound fractions were determined by equilibrium dialysis. Immunohistochemistry was performed to assess levels of pAURKA (T288), phospho-Histone H3 (pHH3), and cleaved caspase 3 (CC3). For survival studies, mice with intracranial tumors were randomized to four cohorts – vehicle, radiotherapy, LY3295668 monotherapy, and LY3295668 plus radiotherapy. The median unbound concentration of LY3295668 was 270.88 nmol/L and 22.33 nmol/kg in plasma and tumor tissue, respectively – significantly higher than the biochemical IC50 of LY3295668 for AURKA inhibition (0.8 nM). A decrease in pHH3(+) cells (0.8% vs. 6.4%, p=0.036) indicated drug-induced mitotic arrest and was accompanied by an increase in CC3(+) cells (6.4% vs. 8.0%, p=0.67). Combination of LY3295668 with radiotherapy prolonged survival compared to either therapy alone in orthotopic GBM PDX models. LY3295668 is well tolerated, achieves pharmacologically-relevant unbound concentrations in GBM PDX models, and is associated with significant target modulation. Preclinical combination of LY3295668 with radiation therapy leads to synergistic effects and supports future clinical study of this multimodal strategy in glioblastoma patients.

Patient Selection Criteria for Deep Brain Stimulation for Dystonia

2020 ◽  
pp. 175-184
Author(s):  
Laura S. Surillo Dahdah ◽  
Rasheda El-Nazer ◽  
Richard B. Dewey ◽  
Padraig O’Suilleabhain ◽  
Shilpa Chitnis
Keyword(s):  
Botulinum Toxin ◽  
Patient Selection ◽  
Muscle Activation ◽  
Age At Onset ◽  
Voluntary Action ◽  
Body Distribution ◽  
Segmental Dystonia ◽  
Generalized Dystonia ◽  
Patient Selection Criteria ◽  
Botulinum Toxin Injections

Dystonia is defined as a movement disorder characterized by sustained or intermittent muscle contractions causing abnormal, often repetitive, movements, postures, or both. Dystonic movements are typically patterned and twisting and may be tremulous. Dystonia is often initiated or worsened by voluntary action and associated with overflow muscle activation. A recent revision now classifies dystonia into two axes: (1) clinical characteristics (age at onset, temporal pattern, body distribution, whether focal, segmental, or generalized; and associated features) and (2) etiology, whether idiopathic/genetic or secondary to other neurological/medical diseases. Pharmacological treatments for dystonia remain generally unsatisfactory and consist of various combinations of levodopa, anticholinergics, muscle-relaxing drugs as well as botulinum toxin injections in focal and segmental dystonia. Overall in outcomes are poor because of limited efficacy and the potential for significant side effects such as sedation and cognitive impairment. A humanitarian-device exemption from the Food and Drug Administration was issued for the treatment of medically refractory symptoms of generalized dystonia with the use of DBS. Bilateral GPi DBS surgery is effective for both generalized and focal dystonia including cervical dystonia and tardive dystonia. DBS may be the best available treatment for disabling symptoms of generalized, cervical, tardive, and other dystonia that have failed to respond to oral drugs and botulinum toxin injections (when applicable) as long as contractures have not developed, because in this situation, DBS will be ineffective. Rigorous patient selection and careful management of comorbidities are essential for favorable outcomes.

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