A study on the CD<Subscript>4</Subscript><Superscript>+</Superscript> CD<Subscript>25</Subscript><Superscript>+</Superscript> regulatory T cells in patients with gastrointestinal malignancies

Objectives Regulatory T cells play an active role in the maintenance of the immune system’s tolerance of both foreign and self antigens. Particularly, CD₄ ⁺ CD₂₅ ⁺ regulatory T cells participate in tumor immunity. The study provided further evidence on the involvement of CD₄ ⁺ CD₂₅ ⁺ regulatory T cells in immune system impairment in patients with gastrointestinal malignancies. Methods Using flow cytometry, CD₄ ⁺ CD₂₅ ⁺ regulatory T cells were analyzed in peripheral blood from 114 patients with gastrointestinal malignancies and 15 healthy controls. Results The prevalence of the CD₂₅ ⁺ subset in CD₄ ⁺ T cells was increased in patients with colorectal carcinoma compared with healthy controls. The phenotic characteristics of the CD₄ ⁺ CD₂₅ ⁺ T cells in patient with malignancies were low expression of CD₄₅ RA and no expression of CD₆₉. Our results indicated that when compared with healthy control, the proportions of CD₄ ⁺ CD₂₅ ⁺ T cells in the peripheral blood of patients with colorectal, gastric, and esophageal carcinoma were significantly higher (P < 0.05) in colorectal carcinoma (22.11 ± 9.65%), gastric carcinoma (17.74 ± 4.24%), and esophageal carcinoma (24.37 ± 4.82)%, respectively. Further analysis on the proportion of CD₄ ⁺ CD₂₅ ⁺ T cells revealed that those patients with gastrointestinal malignancies in stages IV were higher than those of in stage I–III, though no significant difference was observed (P > 0.05). However, the proportion of CD₄ ⁺ CD₂₅ ⁺ T cells in the patients with relapse gastric carcinoma (23.32 ± 4.98%) was significantly higher than that of patients with primary gastric carcinoma (P < 0.01). Conclusions The increased CD₄ ⁺ CD₂₅ ⁺ T cells in patients with gastrointestinal malignancies may be related to immunosuppression and tumor progression. This suggests that elimination or reduction of CD₄ ⁺ CD₂₅ ⁺ regulatory T cells can improve effective tumor immunity for immunotherapy.     

TGFβ<Subscript>1</Subscript> signaling via α<Subscript>V</Subscript>β<Subscript>6</Subscript> integrin

Background

Transforming growth factor β₁ (TGFβ₁) is a potent inhibitor of epithelial cell growth, thus playing an important role in tissue homeostasis. Most carcinoma cells exhibit a reduced sensitivity for TGFβ₁ mediated growth inhibition, suggesting TGFβ₁ participation in the development of these cancers. The tumor suppresor gene DPC4/SMAD4, which is frequently inactivated in carcinoma cells, has been described as a key player in TGFβ₁ mediated growth inhibition. However, some carcinoma cells lacking functional SMAD4 are sensitive to TGFβ₁ induced growth inhibition, thus requiring a SMAD4 independent TGFβ₁ pathway.

Results

Here we report that mature TGFβ₁ is a ligand for the integrin α_Vβ₆, independent of the common integrin binding sequence motif RGD. After TGFβ₁ binds to α_Vβ₆ integrin, different signaling proteins are activated in TGFβ₁-sensitive carcinoma cells, but not in cells that are insensitive to TGFβ₁. Among others, interaction of TGFβ₁ with the α_Vβ₆ integrin resulted in an upregulation of the cell cycle inhibitors p21/WAF1 and p27 leading to growth inhibition in SMAD4 deleted as well as in SMAD4 wildtype carcinoma cells.

Conclusions

Our data provide support for the existence of an alternate TGFβ₁ signaling pathway that is independent of the known SMAD pathway. This alternate pathway involves α_Vβ₆ integrin and the Ras/MAP kinase pathway and does not employ an RGD motif in TGFβ₁-sensitive tumor cells. The combined action of these two pathways seems to be necessary to elicit a complete TGFβ₁ signal.

     

TGFβ<Subscript>1</Subscript> activates c-Jun and Erk1 via α<Subscript>V</Subscript>β<Subscript>6</Subscript> integrin

Transforming growth factor β (TGFβ) plays an important role in animal development and many cellular processes. A variety of cellular functions that are required for tumor metastasis are controlled by integrins, a family of cell adhesion receptors. Overexpression of α_Vβ₆ integrin is associated with lymph node metastasis of gastric carcinomas. It has been demonstrated that a full TGFβ₁ signal requires both α_Vβ₆ integrin and SMAD pathway. TGFβ₁ binds to α_Vβ₆ via the DLXXL motif, a freely accessible amino acid sequence in the mature form of TGFβ₁. Binding of mature TGFβ₁ to α_Vβ₆ leads to immobilization and tyrosine phosphorylation of proteins, which are associated with focal adhesions, a hallmark of integrin-mediated signal transduction. Here, we show that binding of mature TGFβ₁ recruits the mitogen-activated protein kinase kinase kinase 1 (MEKK1), a mediator of c-Jun activation, and the extracellular signaling-regulated kinase-1 (Erk1) to focal adhesions. In addition, the p21-activated kinase 1 (PAK1) is associated with focal adhesions and differentially phosphorylated upon TGFβ₁ stimulation. We conclude that TGFβ₁ activates c-Jun via the MEKK1/p38 MAP kinase pathway and influences cytoskeletal organization. These finding may provide a link between TGFβ₁ and the metastatic behavior of cancers.     

Are <Emphasis Type="Italic">BRCA1</Emphasis>- and <Emphasis Type="Italic">BRCA2</Emphasis>-related breast cancers associated with increased mortality?

There has been contradictory evidence as to whether BRCA1 associated breast cancers have a poorer prognosis than non-BRCA1 cancers. In this issue of Breast Cancer Research Robson and colleagues provide further evidence for poorer survival in BRCA1 carriers and show that it could be attributed to failure to treat small node-negative grade 3 breast cancers with chemotherapy. There still remains little evidence for a survival difference for BRCA2 related breast cancers. Although the high contralateral breast cancer risk is confirmed by this study there is no real evidence for an increase in ipsilateral recurrence or new primary breast cancers in mutation carriers up to the 10-year point.     

Association of genetic polymorphisms of <Emphasis Type="Italic">ER-α</Emphasis> and the estradiol-synthesizing enzyme genes <Emphasis Type="Italic">CYP17</Emphasis> and <Emphasis Type="Italic">CYP19</Emphasis> with breast cancer risk in Chinese women

Estrogen plays a role in breast cancer development, and genetic polymorphisms in estrogen receptor gene ER-α and genes regulating estrogen biosynthesis and metabolisms are associated with the risk of breast cancer in women in western countries. Therefore, we hypothesized that SNPs in ER-α and other estrogen-metabolizing genes contribute to breast cancer risk in Chinese women. In this study, we genotyped polymorphisms in the regulatory regions of ER-α (rs3798577) and other two estrogen-metabolizing enzyme genes CYP17 (rs743572) and CYP19 (rs10046) among 300 breast cancer cases and 390 controls in a Chinese population. Crude and adjusted odds ratios (aORs) and 95% confidence intervals (CIs) were calculated by unconditional logistic regression analyses to estimate breast cancer risk associated with these polymorphisms. We found that the T allele frequency of ER-α was significantly higher in cases (59.8%) than controls (54.5%) (P = 0.047), but no significant difference was found in the genotype distribution. However, postmenopausal breast cancer risk was associated with the CYP17 TC genotype (aOR = 1.77, 95% CI = 1.11–2.83) compared with the TT genotype. The CYP19 variant TC + TT genotypes were associated with both overall cancer risk (TT + TC vs. TT aOR = 1.73, 95% CI = 1.13–2.65) and premenopausal cancer risk (TT + TC vs. TT aOR = 1.78, 95% CI = 1.03–3.09), particularly for ER +/PR + tumors. Furthermore, there were joint effects between CYP19 T and ER-α T varint genotypes (aOR = 1.67, 95% CI = 1.03–2.69 for CYP19 TC + TT vs. CC among ER-α T variant carriers) and between CYP19 T and CYP17 C variant genotypes (aOR = 1.77, 95% CI = 1.11–2.83 for CYP19 TC + TT vs. CC among CYP17 variant C carriers). This study provides evidence that polymorphisms CYP17 rs743572, CYP19 rs10046 and ER-α rs3798577 are associated with breast cancer risk among Chinese women.     

Prolonged and severe thrombocytopenia with pancytopenia induced by radiation-combined temozolomide therapy in a patient with newly diagnosed glioblastoma—analysis of <Emphasis Type="Italic">O</Emphasis><Superscript>6</Superscript>-methylguanine-DNA methyltransferase status

BACKGROUND: We reviewed the risk of second tumor (ST), both malignant and benign, in germinoma survivors followed at the Johns Hopkins Hospital (JHH). METHODS: Between 1977 and 2002, 27 patients with intracranial germinoma were treated with radiation therapy (RT). In the presence of competing events, a cumulative incidence function of ST was estimated using the minimal time interval from the date of diagnosis to the date of ST, date of death, or date of last follow-up. RESULTS: Five patients (18%) developed a ST of which 4 (15%) were malignant. One developed a benign falcine meningioma. The cumulative incidence of ST was 9% at 11 years (95% CI, 0-22%). CONCLUSIONS: The relative contributions of RT and patient susceptibility to a ST cannot be determined but suggests the need for long-term surveillance, including testicular self-exams in male germinoma survivors. Current trials of chemotherapy and reduced RT dose and volume offer the prospect of a lower risk of treatment-induced ST.     

Sex Ratio Distortion in Offspring of Families with <Emphasis Type="BoldItalic">BRCA1</Emphasis> or <Emphasis Type="BoldItalic">BRCA2</Emphasis> Mutant Alleles: An Ascertainment Bias Phenomenon?

Summary Background. There has been controversy regarding whether BRCA1 germline mutations favor female births or whether the sex imbalances observed are attributable to ascertainment bias. Our aims were to compare the sex ratios among offspring of BRCA1-positive, BRCA2-positive, and BRCA-negative families undergoing genetic testing in clinical programs, and to determine whether ascertainment bias is responsible for the observed preponderance of female offspring.Patients and methods. A total of 145 breast and/or ovarian cancer families with mutations in BRCA1 (n = 83) or BRCA2 (n = 62), and 90 families without identifiable mutation were collected for the study from familial cancer clinics in Barcelona, Spain, and Boston, US. Sex ratio was analyzed among all births in the families and offspring of all (tested and obligate) carriers. In order to minimize the effect of family history of cancer, the analysis was also performed among offspring of the most recent generation of mutation-positive carriers who did not have affected children and compared with a control group comprised of the offspring of the most recent adult generation of non-carriers from families with a known mutation.Results. There was a statistically higher proportion of female births in all groups (BRCA1 59% (95% CI = 57–61%), BRCA2 58% (56–61%), and BRCA-negative 59% (56–61%), respectively). The female preponderance persisted in analyses limited to offspring of BRCA1 and BRCA2 carriers (61% (57–65%), and 62% (58–66%), respectively), with no differences between the two mutation groups. In contrast, the excess of female offspring disappeared when ascertainment or recall biases were minimized, 44% (37–52%), and 39% (26–53%) for BRCA1; 51% (44–58%), and 46% (33–60%) for BRCA2.Conclusions. Our findings suggest that there is no asymmetry in birth outcomes among BRCA1 or BRCA2 mutations carriers. Rather ascertainment bias in families participating in genetic testing, or in the family history information they provide is likely to account for excess of female offspring previously reported.     

Diagnostic accuracy of <Superscript>18</Superscript>F–FDG PET/CT and MR imaging in patients with adenoid cystic carcinoma

Background

The aim of this study was to evaluate the value of 18F–FDG PET/CT (PET/CT) and MRI for local and/or whole-body restaging of adenoid cystic carcinoma of the head and neck (ACC).

Methods

Thirty-six patients with ACC underwent conventional MRI of the head and neck and a whole-body PET/CT and were analysed with regards to detection of a local tumor recurrence, lymph node or distant metastases. A consensus interpretation of all available imaging data was used as reference standard. Sensitivity, specificity, diagnostic accuracy, positive and negative predictive values were calculated for MRI and PET/CT.

Results

The sensitivity of PET/CT and MRI was 96% (89%), specificity 89% (89%), PPV 96% (96%), NPV 89% (73%) and accuracy 94% (89%) for detection of local tumors. Additionally, PET/CT revealed lymph node metastases in one patient and distant metastases in 9/36 patients. In three patients secondary primaries were found.

Conclusions

Whole-body PET/CT in addition to MRI of the head and neck improves detection of local tumour and metastastic spread in ACC.

     

<Emphasis Type="Italic">NRAS</Emphasis><Superscript><Emphasis Type="Italic">Q61K</Emphasis></Superscript> mutated diffuse leptomeningeal melanomatosis in an adult patient with a brief review of the so-called “forme fruste” of neurocutaneous melanosis

Primary melanocytic tumors of central nervous system represent rare tumors arising from melanocytes of the leptomeninges. These neoplasms include focal forms like melanocytoma and primary malignant melanoma and diffuse forms like leptomeningeal melanocytosis and primary leptomeningeal melanomatosis. The clinical diagnosis remains challenging, with clinical and radiologic features overlapping with other more common diseases. Here we present a case of a 38 years old male with primary diffuse leptomeningeal melanomatosis with presence of a NRAS^Q61K mutation without features of neurocutaneous melanosis.     

A tandem duplication of <Emphasis Type="Italic">BRCA1</Emphasis> exons 1–19 through <Emphasis Type="Italic">DHX8</Emphasis> exon 2 in four families with hereditary breast and ovarian cancer syndrome

Background

Physical activity is inversely associated with the risk of breast cancer among women in the general population. It is not clear whether or not physical activity is associated with the risk of BRCA-associated breast cancer.

Methods

We conducted a case–control study of 443 matched pairs of BRCA mutation carriers to evaluate the association between physical activity and breast cancer risk. Moderate and vigorous physical activities at ages 12–13, ages 14–17, ages 18–22, ages 23–29 and ages 30–34 were determined using the Nurses’ Health Study II Physical Activity Questionnaire. We estimated mean metabolic equivalent task hours/week for moderate, vigorous and total physical activities overall (ages 12–34), during adolescence (ages 12–17) and during early adulthood (ages 18–34). Logistic regression analysis was used to estimate the odds ratios (OR) and 95% confidence intervals (CI) for total, moderate and strenuous recreational physical activities and breast cancer risk, by menopausal status.

Results

Overall, there was no significant association between total physical activity and subsequent breast cancer risk (OR_{Q4 vs. Q1} = 1.01, 95% CI 0.69–1.47; P-trend = 0.72). Moderate physical activity between ages 12–17 was associated with a 38% decreased risk of premenopausal breast cancer (OR_{Q4 vs. Q1} = 0.62; 95% CI 0.40–0.96; P-trend = 0.01). We found no association between exercise and breast cancer diagnosed after menopause.

Conclusions

These findings suggest that early-life physical activity is associated with a reduced risk of premenopausal breast cancer among BRCA mutation carriers.

Impact

Future prospective analyses, complemented by mechanistic evidence, are warranted in this high-risk population.

     

<Emphasis Type="Italic">TP53</Emphasis> and <Emphasis Type="Italic">CDKN1A</Emphasis> mutation analysis in families with Li–Fraumeni and Li–Fraumeni like syndromes

Li–Fraumeni and Li–Fraumeni like syndromes (LFS/LFL) represent rare cancer–prone conditions associated mostly with sarcomas, breast cancer, brain tumors, and adrenocortical carcinomas. TP53 germline mutations are present in up to 80 % of families with classic Li–Fraumeni syndrome, and in 20–60 % of families with Li–Fraumeni like phenotypes. The frequency of LFS/LFL families with no TP53 mutations detected suggests the involvement of other genes in the syndrome. In this study, we searched for mutations in TP53 in 39 probands from families with criteria for LFS/LFL. We also searched for mutations in the gene encoding the main mediator of p53 in cell cycle arrest, CDKN1A/p21, in all patients with no mutations in TP53. Eight probands carried germline disease-causing mutations in TP53: six missense mutations and two partial gene deletions. No mutations in CDKN1A coding region were detected. TP53 partial deletions in our cohort represented 25 % (2/8) of the mutations found, a much higher frequency than usually reported, emphasizing the need to search for TP53 rearrangements in patients with LFS/LFL phenotypes. Two benign tumors were detected in two TP53 mutation carriers: an adrenocortical adenoma and a neurofibroma, which raises a question about the possible implication of TP53 mutations on the development of such lesions.     

Genetic polymorphisms of <Emphasis Type="Italic">TGF-β</Emphasis>1 &; <Emphasis Type="Italic">TNF-β</Emphasis> and breast cancer risk

Objective. The proliferation of malignant breast epithelial cells is regulated by various stimuli including cytokines and growth factors, thus the variants of those genes may modify the breast cancer risk. To evaluate the potential influences of TGF-1 T²⁹C and TNF- A²⁵²G gene polymorphisms on breast cancer risk, a case–control study was conducted in Korea.Methods. Histologically confirmed breast cancer cases (n = 560) and controls (n=509) with no previous history of cancer were recruited from three teaching hospitals in Seoul, Korea. Genotypes were determined by PCR-CTPP (polymerase chain reaction with confronting two-pair primers) method. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated by unconditional logistic regression model adjusting for age, body mass index, education, parity, age at first full-term pregnancy, and family history of breast cancer. Results. The TGF-1 ²⁹C-allele containing genotypes posed an increased risk of breast cancer (OR=1.3, 95% CI=1.02–1.79), especially in postmenopausal women (OR=1.6, 95% CI=1.01–2.44). Similarly, the TNF- ²⁵²G-allele containing genotypes posed an increased risk of postmenopausal breast cancer (OR=1.7, 95% CI=1.09–2.55). The risk of postmenopausal breast cancer increased in parallel with the number of the risk genotypes (p for trend <0.01). When data were stratified by the presumed non-genetic risk factors, TGF-1 C-allele containing genotypes were found to increase breast cancer risk almost two-fold in postmenopausal women with greater than median body mass index (>22.8 kg/m²) (OR=1.9, 95% CI=1.04–3.37).Conclusion. The results of this study therefore suggest that polymorphisms of TGF-1 and TNF- genes may modify individual susceptibility to breast cancer in Korean women.     

A new <Emphasis Type="Italic">POT1</Emphasis> germline mutation—expanding the spectrum of <Emphasis Type="Italic">POT1</Emphasis>-associated cancers

Melanomas are associated with several hereditary conditions. We present a large family with several family members affected with primary melanomas and dysplastic nevi as well as thyroid cancer and other malignant tumors. Clinical work-up did not reveal a mutation in any of the genes usually considered with evaluation for predisposition to melanoma (BRCA1/2, CDKN2A, CDK4, PTEN, TP53). Whole exome sequencing of five affected family members showed a new variant in POT1. POT1 is associated with the telomere shelterin complex that regulates telomere protection and telomerase access. Germline mutations in POT1 were recently shown to be associated with hereditary predisposition to melanoma. Our findings support a role of POT1 germline mutations in cancer predisposition beyond melanoma development, suggesting a broader phenotype of the POT1-associated tumor predisposition syndrome that might also include thyroid cancer as well as possibly other malignant tumors.     

<Superscript>18</Superscript>F-FDOPA PET and MRI characteristics correlate with degree of malignancy and predict survival in treatment-naïve gliomas: a cross-sectional study

Introduction

To report the potential value of pre-operative ¹⁸F-FDOPA PET and anatomic MRI in diagnosis and prognosis of glioma patients.

Methods

Forty-five patients with a pathological diagnosis of glioma with pre-operative ¹⁸F-FDOPA PET and anatomic MRI were retrospectively examined. The volume of contrast enhancement and T2 hyperintensity on MRI images along with the ratio of maximum ¹⁸F-FDOPA SUV in tumor to normal tissue (T/N SUV_max) were measured and used to predict tumor grade, molecular status, and overall survival (OS).

Results

A significant correlation was observed between WHO grade and: the volume of contrast enhancement (r?=?0.67), volume of T2 hyperintensity (r?=?0.42), and ¹⁸F-FDOPA uptake (r?=?0.60) (P?<?0.01 for each correlation). The volume of contrast enhancement and ¹⁸F-FDOPA T/N SUV_max were significantly higher in glioblastoma (WHO IV) compared with lower grade gliomas (WHO I–III), as well as for high-grade gliomas (WHO III–IV) compared with low-grade gliomas (WHO I–II). Receiver-operator characteristic (ROC) analyses confirmed the volume of contrast enhancement and ¹⁸F-FDOPA T/N SUV_max could each differentiate patient groups. No significant differences in ¹⁸F-FDOPA uptake were observed by IDH or MGMT status. Multivariable Cox regression suggested age (HR 1.16, P?=?0.0001) and continuous measures of ¹⁸F-FDOPA PET T/N SUV_max (HR 4.43, P?=?0.016) were significant prognostic factors for OS in WHO I–IV gliomas.

Conclusions

Current findings suggest a potential role for the use of pre-operative ¹⁸F-FDOPA PET in suspected glioma. Increased ¹⁸F-FDOPA uptake may not only predict higher glioma grade, but also worse OS.

     

15-Deoxy-Δ<Superscript>12,14</Superscript>-prostaglandin J<Subscript>2</Subscript> inhibits G<Subscript>2</Subscript>-M phase progression in human breast cancer cells via the down-regulation of cyclin B1 and survivin expression

The cyclopentenone prostaglandin 15-deoxy-Δ^12,14-prostaglandin J₂ (15d-PGJ₂) exerts a growth inhibitory effect on cancer cells, and this effect is linked to the induction of apoptosis or cell cycle arrest. Induction of apoptosis by 15d-PGJ₂ is associated with the down-regulation of anti-apoptotic proteins. G₀-G₁→S phase progression is inhibited by 15d-PGJ₂ via the degradation of cyclin D1. In this study, we further investigated the mechanism by which 15d-PGJ₂ inhibits cancer cell growth by using the breast cancer cell lines MCF-7 and T-47D. Treatment with 20 μM 15d-PGJ₂ for 72 h completely blocked the growth in both cell lines. However, the proportions of apoptotic MCF-7 and T-47D cells were 21.1% and 40.9%, respectively, indicating that the induction of apoptosis did not appear to fully account for growth inhibition by 15d-PGJ₂. Cell cycle analysis using cells synchronized at the G₀-G₁ or S phase revealed that 15d-PGJ₂ blocked not only G₀-G₁→S phase progression but also G₂-M phase progression. The expression of both cyclins D1 and B1 was decreased by 15d-PGJ₂. Furthermore, 15d-PGJ₂ inhibited aurora-B kinase activity, which coincided with the down-regulation of survivin. Thus, 15d-PGJ₂ induced cell cycle arrest at the G₂-M phase via inhibition of cyclin B1 expression and aurora-B kinase activity. We conclude that survivin may be an important target for 15d-PGJ₂, and its down-regulation may lead to a decrease in aurora-B kinase activity. Naoki Tsuji substantially contributed to this work and should also be considered a first author.     

Association of genetic variations in <Emphasis Type="Italic">RTN4</Emphasis> 3′-UTR with risk for clear cell renal cell carcinoma

Nogo proteins play an important role in the apoptosis of cells, especially in tumor cells. The present study was conducted to evaluate whether the TATC (rs71682980) and CAA (rs34917480) insertion/deletion polymorphisms of RTN4 3′-UTR are associated with clear cell renal cell carcinoma (ccRCC). These two polymorphisms were genotyped in 308 ccRCC patients and 466 healthy controls by polymerase chain reaction polyacrylamide gel electrophoresis (PCR-PAGE). Significantly reduced ccRCC risk was observed to be associated with the TATC_ins/ins genotype carriers (Versus TATC_del/del: adjusted OR 0.53, 95% CI 0.32–0.87, P?=?0.022; Versus TATC_{del/del?del/ins}: adjusted OR 0.57, 95% CI 0.36–0.92, P?=?0.017). After performing stratification analysis, the frequency of TATC_ins/ins genotype was observed to be significantly higher in patients with N0 compared the patients with N1 (P?=?0.003). The present study provide evidence for the first time that the TATC insertion/deletion polymorphism in RTN4 3′-UTR may contributes to ccRCC risk in Chinese Han population.