Response to sunitinib in Chinese KIT-mutated metastatic mucosal melanoma

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. e20017-e20017 ◽  
Author(s):  
Y. Zhu ◽  
L. Si ◽  
Y. Kong ◽  
Z. Chi ◽  
X. Yuan ◽  
...  
Keyword(s):  
Large Scale ◽  
Tissue Sample ◽  
Direct Sequencing ◽  
Kit Mutation ◽  
Over Expression ◽  
Pcr Products ◽  
Grade Ii ◽  
Melanoma Patients ◽  
Exon 11 ◽  
Eye Melanoma

e20017 Background: To investigate the possible relationship between sunitinib response and KIT mutation as well as KIT expression. Methods: We find that a 75-year male patient with KIT-mutated metastatic mucosal (primary: nasal, metastasis: left adrenal gland, lungs and right eye) melanoma has a very good response to sunitinib 37.5mg/d consecutively. We apply IHC sp assay to detect S100, HMB-45, CD117, VEGFR, PDGFR and EGFR in the melanoma sample of this patient. DNA is also extracted from paraffin-embedded melanoma sample of this patient and PCR is carried out to amplify several exons of C-KIT (exon11,13,17,18),B-RAF(exon11, 15) and N- RAS(exon1, 2). PCR products are sent to be directly sequenced. Clinical response, PFS and OS of the patient are evaluated. Results: Strong expression of S100, HMB45, CD117, VEFGR and PDGFR is detected by IHC in the tissue sample of the patient. Furthermore, direct sequencing after PCR reveals that V559A substitution is detected in exon 11 of KIT, with mutation of T into C at 1776, leading to substitution of Valine by Alanine at 559. No mutation was detected in exon 9, 13, 17, 18 of KIT, B-RAF15, 11 or N-RAS1, 2. The patient received sunitinib 37.5mg/d from July 2008 consecutively and evaluated PR (Recist: tumors shrink 70%) lasting for 5 months with Grade II myelosuppression and stomatitis. Now the patient is still alive and received sunitinib. Conclusions: Our case report suggests that response of melanoma patients to sunitinib might be correlated to KIT mutation and expression, and large-scale clinical trial in patients with KIT mutation and over-expression might be useful to shed light on treatment of melanoma patients with individual targeted therapy. No significant financial relationships to disclose.

scholarly journals Cutaneous Hyperpigmentation and Familial Gastrointestinal Stromal Tumor Associated With Germline KIT Mutation Treated With Imatinib: A Chinese Case Report

2020 ◽  
Author(s):  
Wei Yuan ◽  
Wen Huang ◽  
Lei Ren ◽  
Jinghuan Lv ◽  
Chen Xu ◽  
...  
Keyword(s):  
Abdominal Pain ◽  
Gastrointestinal Stromal Tumor ◽  
Mutational Analysis ◽  
Direct Sequencing ◽  
Correct Diagnosis ◽  
Stromal Tumor ◽  
Kit Mutation ◽  
History Of ◽  
Exon 11 ◽  
Cells Of Cajal

Abstract BackgroundFamilial gastrointestinal stromal tumor (GIST) has been identified with multiple GISTs harboring the mutations in germline KIT and PDGFRA. There are only 35 kindreds with germline KIT and 6 with PDGFRA mutations have been reported to date. Familial GIST is often characterized by a series of manifestations, such as multiple lesions, hyperpigmentation, mastocytosis, and dysphagia. Only some kindreds have response to imatinib treatment.Materials and MethodsA 25-year-old Chinese woman presented to the hospital with abdominal pain, and computed tomography (CT) scan showed multiple tumors in the small intestine. Her father had a history of multifocal GISTs, and referred to the hospital with abdominal pain and tumor recurrences last year. Immuhistochemical analysis of CD117 and DOG-1 were performed on tumor samples from the two patients, while KIT mutational analysis was carried out by direct sequencing on DNA from paraffin-embedded specimens and saliva sample.ResultsMultiple GISTs associated with diffuse interstitial cells of Cajal (ICC) hyperplasia were illustrated in these two patients. These tumors were positive for CD117 and DOG-1. The germline mutation at codon 560 of exon 11 (p.V560G) of the KIT gene were found. Treatment with imatinib resulted in favorable responses in both tumor and cutaneous hyperpigmentation.ConclusionsIt is difficult to make a correct diagnosis of familial GIST at first time due to its rarity. This case was finally diagnosed as familial GIST depending on the combination of diffuse ICC hyperplasia, germline KIT mutation, hyperpigmentation and its family history.

scholarly journals Cutaneous Hyperpigmentation and Familial Gastrointestinal Stromal Tumor Associated with Germline KIT Mutation Treated with Imatinib

2020 ◽  
Author(s):  
Wei Yuan ◽  
Wen Huang ◽  
Lei Ren ◽  
Chen Xu ◽  
Lijuan Luan ◽  
...  
Keyword(s):  
Abdominal Pain ◽  
Gastrointestinal Stromal Tumor ◽  
Mutational Analysis ◽  
Direct Sequencing ◽  
Correct Diagnosis ◽  
Saliva Sample ◽  
Stromal Tumor ◽  
Imatinib Treatment ◽  
Kit Mutation ◽  
Exon 11

Abstract Background: Familial gastrointestinal stromal tumor (GIST) has been identified with multiple GISTs containing the mutations in germline KIT and PDGFRA. There are only 35 kindreds with germline KIT and 6 with PDGFRA mutations have been reported so far. Familial GIST is often characterized by a series of manifestations, such as multiple lesions, hyperpigmentation, mastocytosis, and dysphagia. Only some kindreds have response to imatinib treatment.Materials and Methods: A 25-year-old Chinese woman went to hospital because of the abdominal pain, and through the computed tomography (CT) scan showed us the multiple tumors in the small intestine. Her father had a history of multifocal GISTs, and referred to the hospital with abdominal pain and tumor recurrences last year. Immuhistochemical analysis of CD117 and DOG-1 were performed on tumor samples from the two patients, while KIT mutational analysis was carried out by direct sequencing on DNA from paraffin-embedded specimens and saliva sample.Results: Multiple GISTs associated with diffuse interstitial cells of Cajal (ICC) hyperplasia were illustrated in these two patients. These tumors were positive for CD117 and DOG-1. The germline mutation at codon 560 of exon 11 (p.V560G) of the KIT gene were found. The treatment with imatinib resulted in favorable responses in both tumor and cutaneous hyperpigmentation.Conclusions: It is difficult to make a correct diagnosis of familial GIST at first time for its rarity. This case was finally diagnosed as familial GIST depending on the combination of diffuse ICC hyperplasia, germline KIT mutation, hyperpigmentation and its family history.

scholarly journals Detection of erythromycin-resistant determinants by PCR.

1996 ◽  
Vol 40 (11) ◽  
pp. 2562-2566 ◽  
Author(s):  
J Sutcliffe ◽  
T Grebe ◽  
A Tait-Kamradt ◽  
L Wondrack
Keyword(s):  
Direct Sequencing ◽  
Pcr Primers ◽  
Efflux Pumps ◽  
Clinical Isolates ◽  
Gene Products ◽  
Mechanisms Of Resistance ◽  
Erythromycin Resistance ◽  
Surveillance Studies ◽  
Resistance Determinants ◽  
Pcr Products

Erythromycin resistance determinants include Erm methylases, efflux pumps, and inactivating enzymes. To distinguish the different mechanisms of resistance in clinical isolates, PCR primers were designed so that amplification of the partial gene products could be detected in multiplex PCRs. This methodology enables the direct sequencing of amplified PCR products that can be used to compare resistance determinants in clinical strains. Further, this methodology could be useful in surveillance studies of erythromycin-resistant determinants.

scholarly journals A large-scale resource for tissue-specific CRISPR mutagenesis in Drosophila

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Fillip Port ◽  
Claudia Strein ◽  
Mona Stricker ◽  
Benedikt Rauscher ◽  
Florian Heigwer ◽  
...  
Keyword(s):  
Large Scale ◽  
Target Genes ◽  
High Efficiency ◽  
Direct Sequencing ◽  
Fine Tuning ◽  
Genetic Screens ◽  
Efficient Gene ◽  
Crispr Screen ◽  
High Gene ◽  
Multicellular Organisms

Genetic screens are powerful tools for the functional annotation of genomes. In the context of multicellular organisms, interrogation of gene function is greatly facilitated by methods that allow spatial and temporal control of gene abrogation. Here, we describe a large-scale transgenic short guide (sg) RNA library for efficient CRISPR-based disruption of specific target genes in a constitutive or conditional manner. The library consists currently of more than 2600 plasmids and 1700 fly lines with a focus on targeting kinases, phosphatases and transcription factors, each expressing two sgRNAs under control of the Gal4/UAS system. We show that conditional CRISPR mutagenesis is robust across many target genes and can be efficiently employed in various somatic tissues, as well as the germline. In order to prevent artefacts commonly associated with excessive amounts of Cas9 protein, we have developed a series of novel UAS-Cas9 transgenes, which allow fine tuning of Cas9 expression to achieve high gene editing activity without detectable toxicity. Functional assays, as well as direct sequencing of genomic sgRNA target sites, indicates that the vast majority of transgenic sgRNA lines mediate efficient gene disruption. Furthermore, we conducted the so far largest fully transgenic CRISPR screen in any metazoan organism, which further supported the high efficiency and accuracy of our library and revealed many so far uncharacterized genes essential for development.

scholarly journals Human Rhinovirus Molecular Epidemiology and Genotyping in Iranian Military Trainees with Acute Respiratory Symptoms

2021 ◽  
Vol 8 (4) ◽  
Author(s):  
Fahimeh Safarnezhad Tameshkel ◽  
Ali Salimi Jeda ◽  
Ahmad Tavakoli ◽  
Mohammad Hadi Karbalaie Niya ◽  
Morteza Izadi ◽  
...  
Keyword(s):  
Molecular Epidemiology ◽  
Clinical Symptoms ◽  
Respiratory Infections ◽  
Direct Sequencing ◽  
Human Rhinovirus ◽  
P Value ◽  
Rt Pcr ◽  
Respiratory Problems ◽  
Pcr Products ◽  
Tract Infections

Background: Human rhinovirus (HRV) is still the most prevalent viral infection in humans and a significant cause of acute respiratory tract infections (ARTIs) in many communities, including military personnel undergoing basic training. Objectives: In this research, we assessed the molecular epidemiology, genotyping, and phylogenetic classification of HRVs in Iranian military trainees with respiratory infections (RI). Methods: For HRV identification and genotyping, respiratory specimens were obtained, and RT-PCR was conducted for genotyping and phylogenetic analysis of HRV utilizing primers for the 5-UTR region. Results: Among 400 Iranian military trainees (average age of 21 ± 4 years, the range of 18 - 57 years) with respiratory infections, HRV was detected in 29 patients (7%) using RT-PCR. The direct sequencing of PCR products from 10 specimens showed that the incidence of type A (n = 5, 50%) was higher than that of type B (n = 4, 40%) and type C (n = 1, 10%). There were no significant associations between HRV and respiratory and clinical symptoms, blood group, and indoor or outdoor conditions (P-value > 0.05). Conclusions: This research was the first to record HRV as a significant cause of respiratory problems among military trainees in Iran, with a frequency of 7%. The most prevalent genotype was HRV-A, which may be applicable in epidemiological and clinical studies, as well as vaccination plans.

scholarly journals Neoadjuvant treatment of a duodenal GIST revealed a new imatinib-sensitive exon 11 c-KIT-mutation

2016 ◽  
Vol 27 ◽  
pp. iv100
Author(s):  
V. Perfetti ◽  
S. Delfanti ◽  
L. Pugliese ◽  
R. Riboni ◽  
E. Dallera ◽  
...  
Keyword(s):  
Neoadjuvant Treatment ◽  
Kit Mutation ◽  
Duodenal Gist ◽  
Exon 11

scholarly journals No Duplicate KRAS Mutation is Identified on the Same Allele in Gastric or Colorectal Cancer Cells with Multiple KRAS Mutations

2007 ◽  
Vol 35 (4) ◽  
pp. 450-457 ◽  
Author(s):  
K Kimura ◽  
T Nagasaka ◽  
N Hoshizima ◽  
H Sasamoto ◽  
K Notohara ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Gastric Cancer ◽  
Cancer Cells ◽  
Kras Mutation ◽  
Direct Sequencing ◽  
Gastric Cancer Cells ◽  
Kras Mutations ◽  
Colorectal Cancer Cells ◽  
Pcr Products ◽  
Codon 12 And 13

Codon 12 and 13 mutations in 170 colorectal cancer (CRC) and 66 gastric cancer (GC) specimens were analysed by an ‘enriched’ polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) method. All identified mutations were verified by direct sequencing of the second PCR products. Among the 170 CRC specimens, mutations were identified in 47 (28%) and 13 (7.6%) cases in codons 12 and 13, respectively. In the 66 GC specimens examined, however, mutations in codons 12 and 13 were only detected in two (3.0%) and one (1.5%) cases, respectively. Mutations in both codon 12 and 13 were found in 3/170 (1.8%) CRCs and 1/66 (1.5%) GCs. Duplicate mutations were never identified in the same allele, which was confirmed by direct sequencing of the second amplified products. The majority of colorectal and gastric cancer cells with KRAS mutations are homogeneous because they have the same KRAS mutation. A few colorectal or gastric cancers, however, showed heterogeneity, as verified by the fact that single mutations were identified in the same allele.

Direct sequencing of PCR products using the Maxam-Gilbert method

1990 ◽  
Vol 7 (5) ◽  
pp. 142-143 ◽  
Author(s):  
Stefan Stamm ◽  
Frank M. Longo
Keyword(s):  
Direct Sequencing ◽  
Pcr Products
Sign in / Sign up

Export Citation Format

Share Document