Point Mutation In Exon Research Articles

Characteristics of Epidermal Growth Factor Receptor with Rare Mutations in Non-small Cell Lung Cancer and the Effect of EGFR Tyrosine Kinase Inhibitors on Them

背景与目的晚肺腺癌是肺癌中最常见的类型，表皮生长因子受体酪氨酸激酶抑制剂（epidermal growth factor receptor tyrosine kinase inhibitors, EGFR-TKIs）目前已成为EGFR突变型非小细胞肺癌（non-small cell lung cancer, NSCLC）的一线标准治疗。经典型突变（19外显子缺失突变和21外显子L858R突变）应用EGFR-TKIs治疗的效果已有大量研究和报道，而少见或复合突变类型的相关报道较少，具体疗效尚未完全统一定论。方法对2016年8月-2018年4月就诊于郑州大学第一附属医院的150例经基因检测证实为EGFR少见突变的NSCLC患者进行回顾性研究，分析其突变类型及临床病理特征，并对其中48例接受EGFR-TKIs Ⅰ/Ⅱ/Ⅲ线治疗的患者的疗效进行描述和评价。结果将全部150例患者按突变类型分为4组，分别为18外显子G719X突变46例（30.7%）、21外显子L861Q突变45例（30.0%）、其他单一少见突变14例（9.3%）和复合突变45例（30%）。EGFR基因少见或复合突变类型与性别、年龄、分期、病理类型及吸烟史均无关。对于48例接受EGFR-TKIs治疗的患者，4组不同类型突变的患者客观缓解率（objective response rate, ORR）和疾病控制率（disease control rate, DCR）差异无统计学意义（54.5% vs 30.0% vs 0.0% vs 35.7%, χ2=3.200, P=0.34; 90.9% vs 85.0% vs 66.7% vs 92.9%, χ2=2.162, P=0.59）。中位无进展生存期（median progress free survival, mPFS）为11.0个月（95%CI: 4.4-17.6），在EGFR基因突变不同类型分组中分别为[15.8个月（95%CI: 9.5-22.2）、8.0个月（95%CI: 5.1-11.0）、4.9个月（95%CI: 1.4-8.4）、23.1个月（95%CI: 15.8-30.4）]（χ2=7.876, P=0.049）。结论不同类型的EGFR少见或复合突变类型应用EGFR的疗效不尽相同，复合突变组PFS可能优于单一少见突变组。接下来有必要进一步进行大样本量的研究，发现新的敏感靶点和研究新一代的药物对于接受现有治疗效果欠佳的患者也是值得期待的。

A multicenter, open label, randomized phase II study of osimertinib plus ramucirumab versus osimertinib alone as initial chemotherapy for EGFR mutation-positive non-squamous non-small cell lung cancer: TORG1833.

TPS9120 Background: Vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) pathways are shown to be interrelated in several preclinical studies. Furthermore, recent clinical studies have shown the adding effect of an anti VEGF monoclonal antibody with an epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) for the non-small-cell lung cancer (NSCLC) patients with EGFR mutation. Thus, osimertinib plus ramucirumab would be the promising candidate for the new standard treatment in EGFR mutation positive NSCLC. Methods: This study is an investigator initiated trial. Previously untreated EGFR mutation positive advanced non squamous NSCLC patients aged 20 years or older with a performance status of 0 or 1 are randomized at a 1:1 ratio to receive osimertinib (80mg) every day either without or with ramucirumab (10mg/kg) every 2 weeks until evidence of disease progression or development of unacceptable toxicity. The primary endpoint of the study is progression free survival (PFS) assessed by the central image reviewer. Secondly endpoints include PFS (assessed by an attending physician), objective response rate (ORR), disease control rate (DCR), duration of response (DOR), overall survival (OS), safety and toxicity profile. Stratification factors are gender and the type of EGFR mutation (exon 19 deletion, Leu858Arg point mutation in exon 21). We determined that, with a sample size of 120 patients (60 in each arm), the trial will have 80% power to show a hazard ratio for disease progression or death of 0.667 at a one-sided alpha level of 0.2 (as calculated on the basis of 80 such events) for comparison between the two arms with 1.5-year accrual and 2-year follow-up periods. Study enrollment began in November 2018 and is continued for 3.5 years among 20 sites of Thoracic Oncology Research Group (TORG). Seven patients were enrolled at time of submission. Clinical trial information: 184146.

Apparent diffusion coefficient and genetic mutations as effective indexes for radiotherapy time of brain metastases from lung cancer retrospective analysis.

e20641 Background: The potential value of apparent diffusion coefficient (ADC) before chemoradiotherapy and genetic mutation as monitors for radiotherapy time of lung adenocarcinoma patients with brain metastases were retrospective analysed. Methods: Fifty-seven patients of lung adenocarcinoma with brain metastases were enrolled into the clinical trial. Conventional magnetic resonance imaging (MRI) examination and diffusion weighted imaging (DWI) were performed one week before therapy and after two cycles of chemotherapy or after one month targeted therapy. Symptoms such as headache, nausea, vomiting would be alleviated with mannitol and dexamethasone. Chemotherapy,once every 3 weeks, using in these patients with wild gene included pemetrexed, DDP, bevacizumab (pemetrexed 500mg per square meter ; DDP 75mg per square meter and bevacizumab 15mg per kilogram). 27 Patients with genetic mutations includ ed 1 patient with ALK genetic mutations, 2 patients with T790 genetic mutations, the presence of EGFR mutations was determined by the presence of deletions with exons 19 in 9 patients and 20 in 10 patients and L858R point mutations in exon 21 in 5 patients. Patients with genetic mutations were given targeted drug therapy. After these above treatment, if brain metastasis symptoms persist, brain radiotherapy was performed ahead of schedule. Patients with no obvious symptoms or relieved symptoms after targeted therapy or chemotherapy, two-month review was performed with enhanced MRI, radiotherapy was not added until the lesion no longer continues to shrink. Results: In the cohort of patients with wild gene , when ADC values less than 1.01 in 15 patients, the maximum diameter of tumors were significantly decreased after two treatment cycles, While 14 patients with ADC values greater than 1.01, the maximum diameter of the tumors are no difference after two treatment cycles. After targeted therapy for one month, In patients with ALK and T790 genetic mutations, the maximum diameter of tumors were significantly decreased, while no difference in 18 patients and decreased in 6 patients with presence of EGFR. Conclusions: ADC values and genetic mutation may as effective indexes of radiotherapy time for lung adenocarcinoma patients with brain metastases.

Identification of Two Novel Mutations in KDM3A Regulatory Gene in Iranian Infertile Males

Background:KDM3A is a key epigenetic regulator expressed in the testis and is required for packaging and condensation of sperm chromatin. To this point, the association of the KDM3A gene with infertility has not been studied in human. The aim of this study was to screen any new mutation in KDM3A gene to explore more details of human male infertility.Methods:In this work, 150 infertile men (oligozoospermia and azoospermia) and 150 normal healthy fathers were studied. Polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and sequencing were used to screen any mutation in exons 12, 22, and 24 of KDM3A.Results:The infertile men showed various SSCP patterns for the exons 12 and 24, but not for exon 22. A transversion point mutation in exon 12 and a single nucleotide deletion in exon 24 were detected using sequencing analysis. The transversion mutation was located in the preceding exon of lysine-specific demethylase1 and Jumonji (Jmj)-C domain and the later one (deletion) in the cupin-like motif of KDM3A protein. Neither Y chromosome microdeletions nor partial azoospermia factor deletion was found in these patients.Conclusion:The mutations found in infertile men with otherwise unexplained severe spermatogenic failure could be considered as the origin of their abnormalities.

Function and Immunogenicity of Gene-corrected iPSC-derived Hepatocyte-Like Cells in Restoring Low Density Lipoprotein Uptake in Homozygous Familial Hypercholesterolemia

Gene correction of induced pluripotent stem cells (iPSCs) has therapeutic potential for treating homozygous familial hypercholesterolemia (HoFH) associated with low-density lipoprotein (LDL) receptor (LDLR) dysfunction. However, few data exist regarding the functional recovery and immunogenicity of LDLR gene-corrected iPSC-derived hepatocyte-like cells (HLCs) obtained from an HoFH patient. Therefore, we generated iPSC-derived HLCs from an HoFH patient harbouring a point mutation (NM_000527.4:c.901 G > T) in exon 6 of LDLR, and examined their function and immunogenicity. From the patient’s iPSCs, one homozygous gene-corrected HoFH-iPSC clone and two heterozygous clones were generated using the CRISPR/Cas9 method. Both types of iPSC-derived HLCs showed recovery of the function of LDL uptake in immunofluorescence staining analysis. Furthermore, these gene-corrected iPSC-derived HLCs showed little immunogenicity against the patient’s peripheral blood mononuclear cells in a cell-mediated cytotoxicity assay. These results demonstrate that LDL uptake of iPSC-derived HLCs from HoFH can be restored by gene correction without the appearance of further immunogenicity, suggesting that gene-corrected iPSC-derived HLCs are applicable to the treatment of HoFH.

Pneumatosis intestinalis induced by osimertinib in a patient with lung adenocarcinoma harbouring epidermal growth factor receptor gene mutation with simultaneously detected exon 19 deletion and T790\u2009M point mutation: a case report

BackgroundPneumatosis intestinalis is a rare adverse event that occurs in patients with lung cancer, especially those undergoing treatment with epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKI). Osimertinib is the most recently approved EGFR-TKI, and its usage is increasing in clinical practice for lung cancer patients who have mutations in the EGFR gene.Case presentationA 74-year-old woman with clinical stage IV (T2aN2M1b) lung adenocarcinoma was determined to have EGFR gene mutations, namely a deletion in exon 19 and a point mutation (T790 M) in exon 20. Osimertinib was started as seventh-line therapy. Follow-up computed tomography on the 97th day after osimertinib administration incidentally demonstrated intra-mural air in the transverse colon, as well as intrahepatic portal vein gas. Pneumatosis intestinalis and portal vein gas improved by fasting and temporary interruption of osimertinib. Osimertinib was then restarted and continued without recurrence of pneumatosis intestinalis. Overall, following progression-free survival of 12.2 months, with an overall duration of administration of 19.4 months (581 days), osimertinib was continued during beyond-progressive disease status, until a few days before the patient died of lung cancer.ConclusionsPneumatosis intestinalis should be noted as an important adverse event that can occur with administration of osimertinib; thus far, such an event has never been reported. This was a valuable case in which osimertinib was successfully restarted after complete recovery from pneumatosis intestinalis, such that further extended administration of osimertinib was achieved.

Clinical and genetic characteristics of female dystrophinopathy carriers.

The present study aimed to determine the genetic status of manifesting carriers (MCs) of Duchenne muscular dystrophy (DMD)/Becker muscular dystrophy (BMD) and asymptomatic carriers with a family history of DMD/BMD, and identify potential simple and reliable methods for screening dystrophinopathy carriers. Clinical data from probable carriers and MCs were collected and analyzed. MCs underwent multiplex ligation-dependent probe amplification (MLPA) for dystrophin gene exons combined with muscle disease panel test based on a next-generation sequencing (NGS) platform. In addition, the status of probable carriers was determined by MLPA or Sanger sequencing, according to the mutations of probands. A total of 154 female were enrolled, among which 78 cases were found to be carriers, including 4 MCs and 74 asymptomatic female carriers. The 4 MCs exhibited duplication mutations. Among the 74 asymptomatic carriers, 41.89% harbored deletion mutations, including 2 cases with suspected germline mosaicism and no mutation in the dystrophin gene, while 44.59% harbored point mutations in exons and only 10 cases (13.51%) carried duplication mutations. The area under the receiver operating characteristic (ROC) curve of creatine kinase (CK) was 0.822, with a sensitivity of 65.38% and specificity of 92.1%. In addition, DMD was positively correlated with the CK, alanine transaminase and aspartate transaminase levels of the carriers. MLPA for exons of the dystrophin gene, along with NGS and Sanger sequencing, was effective for the diagnosis of MCs and for determining the status of probable carriers. The ROC curve analysis also demonstrated that CK level was an excellent predictor for distinguishing DMD/BMD carriers.

A novel homozygous point mutation and deletion in exon 3 of growth hormone receptor causes laron syndrome: A case study

A novel homozygous point mutation and deletion in exon 3 of growth hormone receptor causes laron syndrome: A case study

Generation of 3 human induced pluripotent stem cell lines LUMCi005-A, B and C from a Hereditary Cerebral Hemorrhage with Amyloidosis-Dutch type patient

Hereditary Cerebral Hemorrhage with Amyloidosis-Dutch type (HCHWA-D) is an autosomal dominant hereditary disease caused by a point mutation in exon 17 of the APP gene. We generated human induced pluripotent stem cells (hiPSCs) from a symptomatic HCHWA-D patient by using non-integrating Sendai virus (SeV). The newly generated hiPSCs express all pluripotency markers, have a normal karyotype, carry the Dutch mutation, can differentiate in the three germ layers in vitro and are SeV free.

Breast cancer and synchronous multiple primary lung adenocarcinomas with heterogeneous mutations: a case report

BackgroundMultiple primary malignant tumors (MPMT) refers to the presence of two or more primary cancers of different organs in the same patient. MPMT is a sparse disease in the past, but there has been a gradual increase in the morbidity. Since multiple primary malignant tumors treatment methods differ, it is essential for clinicians to be able to distinguish between separate primary lesions and metastasis.Case presentationWe present the case of a 57-year-old woman with MPMT presenting with cancer in the left breast and synchronous double primary lung adenocarcinomas. We used IHC and epidermal growth factor receptor(EGFR)mutation to analyze genomic alteration profiles in the patient to validate the difference among the pathological assessments and the clinical differences between double primary lesions of lung and breast. EGFR gene analysis of breast cancer lesion revealed no mutations. The left and right lower lobe lung adenocarcinomas contained EGFR gene mutations: an L858R point mutation in exon 21 in the left lesion and a deletion mutation in exon 19 in the right lesion. The breast cancer and both lung adenocarcinomas were surgically resected. To date, the patient has remained disease-free.ConclusionsBoth pathological and molecular assessment adapted in the current study appeared necessary. Mutational analysis of the EGFR gene provided important information not only in the diagnosis and but also in the treatment of MPMT.

The role of epidermal growth factor receptor (EGFR) common gene mutations in Iranian women with uterine fibroids.

Uterine myomas are benign uterine tumors that originate from smooth muscle cells of the myometrium. This common complication can be associated with irreversible complications, including infertility and malignancy. Better understanding of the genetic characteristics of myoma may effect on treatment. Epidermal growth factor receptor gene (EGFR) is one of the most important genes that has the major role in the pathogenesis of myoma, cell growth, differentiation, proliferation and mutagenesis. The aim of this study was to investigate EGFR common gene mutations in Iranian women with uterine fibroids. In this case-control study, Common EGFR gene mutations in exons 21 and exons 19 of 100 women with uterine leiomyoma as cases and 100 healthy women as controls were studied. To investigate deletion mutations of exon 19 (rs121913438) and point mutations of exon 21 (rs121434568), Tetra ARMS/PCR, ARMS and conventional PCR methods were used respectively and the results were analyzed using χ 2 test. Odds ratios (ORs) and 95% confidence intervals (CI) were estimated using logistic regression with control for age. Our results showed significant difference in genotypes frequency of (TT, TG, GG) for exon 21 and (WW, WD, DD) of exon 19 among cases and controls (P-Value = 5.672e-20) and (P-Value = 3.242e-15). There was a significant relationship between [G] allele and risk uterine myoma (P-Value = 3.018e-36) and the presence of [G] allele increased the chance of developing the disease OR = 0.004, 95% CI 0.001-0.013. The result also showed significant relationship between [D] allele and risk of uterine myoma (P-Value = 1.324e-15). In addition, presence of [D] allele, increased the chance of developing the disease (OR = 0.008, 95% C.I. = 0.002-0.033). The results indicated a significant correlation between mutations in exon 19 (rs121913438) and exon 21(rs121434568) of EGFR gene and susceptibility of myoma in the study population.

TAS6417, A Novel EGFR Inhibitor Targeting Exon 20 Insertion Mutations.

Activating mutations in the EGFR gene are important targets in cancer therapy because they are key drivers of non-small cell lung cancer (NSCLC). Although almost all common EGFR mutations, such as exon 19 deletions and the L858R point mutation in exon 21, are sensitive to EGFR-tyrosine kinase inhibitor (TKI) therapies, NSCLC driven by EGFR exon 20 insertion mutations is associated with poor clinical outcomes due to dose-limiting toxicity, demonstrating the need for a novel therapy. TAS6417 is a novel EGFR inhibitor that targets EGFR exon 20 insertion mutations while sparing wild-type (WT) EGFR. In cell viability assays using Ba/F3 cells engineered to express human EGFR, TAS6417 inhibited EGFR with various exon 20 insertion mutations more potently than it inhibited the WT. Western blot analysis revealed that TAS6417 inhibited EGFR phosphorylation and downstream molecules in NSCLC cell lines expressing EGFR exon 20 insertions, resulting in caspase activation. These characteristics led to marked tumor regression in vivo in both a genetically engineered model and in a patient-derived xenograft model. Furthermore, TAS6417 provided a survival benefit with good tolerability in a lung orthotopic implantation mouse model. These findings support the clinical evaluation of TAS6417 as an efficacious drug candidate for patients with NSCLC harboring EGFR exon 20 insertion mutations. Mol Cancer Ther; 17(8); 1648-58. ©2018 AACR.

First-generation EGFR tyrosine kinase inhibitor therapy in 106 patients with compound EGFR-mutated lung cancer: a single institution\u2019s clinical practice experience

BackgroundThe antitumour efficacy of tyrosine kinase inhibitors (TKIs) in lung cancer patients with compound epidermal growth factor receptor (EGFR) mutations has not been resolved. Our study summarizes a single institutional experience of first-generation TKI therapy for lung cancers with compound EGFR mutations.MethodsA total of 106 consecutive patients with tumours bearing compound EGFR mutations were identified between January 2012 and May 2016; all patients received first-generation TKI therapy. Deletions in exon 19 and the L858R point mutation in exon 21 were considered common mutations; T790M was considered separately because of its association with TKIs resistances. Any other mutation was defined as a rare mutation. Patients were divided as follows: double common mutations (group A); common plus T790M mutations (group B); common plus rare mutations (group C); double rare mutations (group D); and rare plus T790M mutations (group E). A separate group of 115 consecutive patients with a single common mutation was created for comparative analysis (group F).ResultsThe frequency of patients with compound EGFR was 2.9% (114/3925) and their response rate to first-generation TKIs was 50.9%, which was not significantly different from group F (67.0%, P = 0.088). The progression-free survival (PFS) of the 106 patients receiving TKI therapy was worse than that of group F (median, 9.1 vs. 13.0 months, respectively; P < 0.001). The PFS of the compound mutation group was shorter than that of the single common mutation group (median, 10.1 months in group A, P = 0.240; 9.1 months in group B, P < 0.001; 9.6 months in group C, P = 0.010; 6.5 months in group D, P = 0.048; 5.4 months in group E, P = 0.017). Patients with a co-occurring mutation in exon 20 (excluding T790M) exhibited significantly worse PFS than the patients with other compound mutations or with a single common mutation (median, 6.5 vs. 9.1 vs. 13.0 months, respectively, P = 0.002).ConclusionsThere was significant heterogeneity among the compound EGFR mutations and their response to first-generation TKIs. Individualized treatment in clinical practice should be considered for each case.

Genetically Engineered iPSC-Derived FTDP-17 MAPT Neurons Display Mutation-Specific Neurodegenerative and Neurodevelopmental Phenotypes.

SummaryTauopathies such as frontotemporal dementia (FTD) remain incurable to date, partially due to the lack of translational in vitro disease models. The MAPT gene, encoding the microtubule-associated protein tau, has been shown to play an important role in FTD pathogenesis. Therefore, we used zinc finger nucleases to introduce two MAPT mutations into healthy donor induced pluripotent stem cells (iPSCs). The IVS10+16 mutation increases the expression of 4R tau, while the P301S mutation is pro-aggregant. Whole-transcriptome analysis of MAPT IVS10+16 neurons reveals neuronal subtype differences, reduced neural progenitor proliferation potential, and aberrant WNT/SHH signaling. Notably, these neurodevelopmental phenotypes could be recapitulated in neurons from patients carrying the MAPT IVS10+16 mutation. Moreover, the additional pro-aggregant P301S mutation revealed additional phenotypes, such as an increased calcium burst frequency, reduced lysosomal acidity, tau oligomerization, and neurodegeneration. This series of iPSCs could serve as a platform to unravel a potential link between pathogenic 4R tau and FTD.

Correlation of plasma erlotinib trough concentration with skin rash in Chinese NSCLC patients harboring exon 19 deletion mutation.

Erlotinib is an essential drug for non-small cell lung cancer (NSCLC) patients harboring epidermal growth factor receptor (EGFR) activating mutations. The relationship between the pharmacokinetics and skin rash and diarrhea of erlotinib in Chinese patients with EGFR mutated NSCLC is unknown. In this study, we evaluated the variability in erlotinib trough concentration and its relationship with the severity of skin rash and diarrhea in patients with two common types of EGFR mutations: a deletion in exon 19 and point mutations in exon 21 L858R. EGFR mutation-positive Chinese patients (n = 52) treated with erlotinib were included in our study; the steady-state trough concentrations were assessed; and the occurrence and severity of skin rash and diarrhea after the onset of treatment with erlotinib were recorded. The patients were divided into two groups by mutation type (exon 19 deletions or exon 21 L858R point mutations). Occurrence and severity of skin rash and diarrhea was analyzed in both groups. The overall mean (± SD) steady-state trough concentration for erlotinib was 1380 ± 663ng/mL, and there was no significant difference of erlotinib concentrations between the two mutation groups. Occurrence and severity of skin rash was significantly associated with trough concentration in patients with exon 19 deletions but not exon 21 L858R point mutations. Significant association of erlotinib concentrations with diarrhea was found neither in the exon 19 deletions group nor in the exon 21 L858R point mutation group. The occurrence and severity of skin rash correlated with increase in erlotinib trough concentrations only in Chinese patients with exon 19 deletion; the erlotinib trough concentrations were not associated with diarrhea.

Genotypes and phenotypes in 20 Chinese patients with type 2 Gaucher disease

BackgroundGaucher disease (GD) is one of the most common lysosomal storage diseases resulting from a deficiency of glucocerebrosidase. Three main types have been described, with type 2 being the most rare and severe form. Here we investigated the clinical symptoms and mutation spectrum in 20 unrelated type 2 GD patients. MethodThe diagnosis of GD was based on the acid β-glucocerebrosidase (GBA) enzyme activity and direct sequencing of the GBA gene. GBA activity was measured in leukocytes and the GBA gene was amplified by a polymerase chain reaction (PCR). For patient 7, the GBA gene was analyzed by PCR as well as quantitative real-time PCR. ResultsThe age of onset was under 12 months for all patients. All patients experienced severe neurological involvement. A total of 19 different GBA gene mutations were identified, including 6 novel mutations: two were exonic point mutations, c.1127T > C (p.Phe376Ser), c.1418T > G (p.Val473Gly); one was splicing error, ISV7-1G > C; one was insertion, c.717_718insACAG; and the other two were a gross deletion that involved exon 6 and a recombinant allele. The most prevalent mutation was Leu483Pro, which constituted 42.5% of all mutant alleles and was associated with a neurological form in Chinese GD patients as calculated by a Fisher’s exact test. ConclusionThe clinical characteristics of Chinese type 2 GD were consistent with reports from other ethnic populations. We identified 6 novel mutations that contribute to the spectrum of GBA gene mutations. Our study confirmed that GD patients with the Leu483Pro allele were prone to experience neurological symptoms.

Gastrointestinal perforation during treatment with erlotinib plus bevacizumab in two patients with non-small cell lung cancer exhibiting epidermal growth factor receptor mutations: A case report.

A previous randomized phase II study in patients with non-small cell lung cancer (NSCLC) identified that combination treatment with erlotinib plus bevacizumab prolonged progression-free survival compared with erlotinib alone. However, combination bevacizumab and erlotinib treatment generally increased the risk of severe adverse events, including hemorrhage, thrombosis, fistula formation and gastrointestinal perforation. The present report describes two patients with NSCLC harboring epidermal growth factor receptor (EGFR) mutations, who experienced gastrointestinal perforation associated with erlotinib plus bevacizumab combination therapy. The first patient, a 67-year-old male with stage IIIB lung adenocarcinoma harboring a L858R point mutation in EGFR exon 21, received concurrent chemoradiotherapy. However, seven months later, the patient experienced a relapse and was administered erlotinib plus bevacizumab treatment. A total of two months subsequent to commencing treatment, the patient developed a perforated duodenal ulcer. The second patient, a 66-year-old male with lung adenocarcinoma harboring a deletion in EGFR exon 19 and multiple pulmonary metastases, demonstrated a partial response to erlotinib plus bevacizumab treatment. A total of seven months subsequent to starting treatment, the patient experienced lower abdominal pain, and abdominal computed tomography confirmed a diagnosis of colocutaneous fistula complicating sigmoid diverticulitis. Following repair of the perforation, both patients were restarted on erlotinib treatment alone. Gastrointestinal perforation may be a potentially severe adverse event of erlotinib plus bevacizumab combination therapy, even in the absence of tumor metastasis in the abdomen.

Identification of factor VIII gene mutations in patients with haemophilia A

This study aimed to identify common FVIII gene mutations in Iraqi patients by molecular analysis and to investigate the correlations between mutations and disease severity. This study included 25 Iraqi patients with haemophilia A, 18 of whom were male haemophilia patients and 7 of whom were carrier mothers, from seven unrelated families. After determining the purity and concentration of the extracted DNA, selected areas of the FVIII gene, including exons 18, 22, 23, and 24 and intron 22, were amplified. Sequencing of all exons and intron 22 was conducted in all patients and controls. Sequencing analysis revealed that many mutations were distributed among the exons (18, 22, 23, and 24) and intron 22; most of the identified mutations were point mutations in exons, predominantly in exon 24. Exon 18 mutations were detected in one male and one mother as a carrier. Exon 22 mutations were detected in four patients, and exon 23 mutations were detected in two males and two carrier mothers. Exon 24 mutations were detected in 12 haemophilia patients and two carrier mothers. Additionally, seven inversion mutations were detected in haemophilia patients. The results also revealed positive correlations between disease severity and mutations in intron 22 and exon 24. We therefore concluded that exon 24 mutations are the most frequent type of mutation that occurs in haemophilia patients in Iraq, followed by intron 22 mutations, and that almost all severe haemophilia cases have mutations located in these two gene segments. Keywords: Camptothecin, Carotenoid, Cellulase enzyme, Extraction, HPLC

Losmapimod Overcomes Gefitinib Resistance in Non-small Cell Lung Cancer by Preventing Tetraploidization

The epidermal growth factor receptor (EGFR) is known to play a critical role in non-small cell lung cancer (NSCLC). Constitutively active EGFR mutations, including in-frame deletion in exon 19 and L858R point mutation in exon 21, contribute about 90% of all EGFR-activating mutations in NSCLC. Although oral EGFR-tyrosine kinase inhibitors (TKIs), gefitinib and erlotinib, show dramatic clinical efficacy with significantly prolonged progression-free survival in patients harboring these EGFR-activating mutations, most of these patients will eventually develop acquired resistance. Researchers have recently named genomic instability as one of the hallmarks of cancer. Genomic instability usually involves a transient phase of polyploidization, in particular tetraploidization. Tetraploid cells can undergo asymmetric cell division or chromosome loss, leading to tumor heterogeneity and multidrug resistance. Therefore, identification of signaling pathways involved in tetraploidization is crucial in overcoming drug resistance. In our present study, we found that gefitinib could activate YAP-MKK3/6-p38 MAPK-STAT3 signaling and induce tetraploidization in gefitinib-resistance cells. Using p38 MAPK inhibitors, SB203580 and losmapimod, we could eliminate gefitinib-induced tetraploidization and overcome gefitinib-resistance. In addition, shRNA approach to knockdown p38α MAPK could prevent tetraploidy formation and showed significant inhibition of cancer cell growth. Finally, in an in vivo study, losmapimod could successfully overcome gefitinib resistance using an in-house established patient-derived xenograft (PDX) mouse model. Overall, these findings suggest that losmapimod could be a potential clinical agent to overcome gefitinib resistance in NSCLC.

Adrenocortical carcinoma characterized by gynecomastia: A case report.

.We present a 4-yr-old boy with adrenocortical carcinoma (ACC), diagnosed due to the appearance of gynecomastia as the presenting symptom. Six months prior to admission, an acute growth spurt along with the development of bilateral breast swelling was observed. He did not present any features of virilization, including enlargement of the testes, increase in testis volume, and penis size. Laboratory investigations showed gonadotropin-independent hypergonadism, with low LH/ FSH levels and elevated estradiol/testosterone levels. Abdominal computed tomography revealed a large heterogeneous mass adjacent to the right kidney and below the liver. Pathological investigations of the biopsy specimen demonstrated that the tumor was an ACC. Pre- and post-operative combination chemotherapy with mitotane was administered and surgical resection was carried out. Post-surgery, the elevated estradiol/testosterone concentrations reverted to within the reference range. Urinary steroid profile and tissue concentration analysis of estradiol and testosterone indicated the presence of estrogen in the ACC tissue. An investigation for TP53 gene aberrations revealed the presence of a germline point mutation in exon 4 (c.215C>G (p.Pro72Arg)). In ACC, the most common symptom is virilization, and feminization, characterized by gynecomastia, is very rare. However, a diagnostic possibility of ACC should be considered when we encounter patients who have developed gynecomastia without the influence of causative factors such as obesity or puberty, and do not present with the typical signs of virilization.