Primary Thymic Lymphoma Research Articles

Thymic lymphoma detection in RORγ knockout mice using 5-hydroxymethylcytosine profiling of circulating cell-free DNA

A high incidence of thymic lymphoma has been noted in mice deficient of retinoid-related orphan receptor γ2 (RORγ2), which is required for differentiation of naïve CD4+ T cells into TH17 cells. Using a RORγ homozygous knockout (KO) mouse model of thymic lymphoma, we characterized this tumor progression and investigated the utility of 5-hydroxymethylcytosine (5hmC) signatures as a non-invasive circulating biomarker for early prediction of malignancy. No evidence for malignancy was noted in the wild-type mice, while primary thymic lymphoma with multi-organ metastasis was observed microscopically in 97% of the homozygous RORγ KO mice. The severity of thymic lymphoma was not age-dependent in the KO mice of 2 to 4 months old. Differential enrichment of 5hmC in thymic DNA and plasma cell-free DNA (cfDNA) was compared across different stages of tumor progression. Random forest modeling of plasma cfDNA achieved good predictivity (AUC = 0.74) in distinguishing early non-metastatic thymic lymphoma compared to cancer-free controls, while perfect predictivity was achieved with advanced multi-organ metastatic disease (AUC = 1.00). Lymphoid-specific genes involved in thymocyte selection during T cell development (Themis, Tox) were differentially enriched in both plasma and thymic tissue. This could help in differentiating thymic lymphoma from other tumors commonly detected in rodent carcinogenicity studies used in pharmaceutical drug development to inform human malignancy risk. Overall, these results provide a proof-of-concept for using circulating cfDNA profiles in rodent carcinogenicity studies for early risk assessment of novel pharmaceutical targets.

Incidence, mortality, and survival analyses of patients with thymic lymphoma.

ObjectivesTo explore the clinical and prognostic characteristics of thymic lymphoma and the effects of current treatments on the prognosis.MethodsPatients diagnosed as primary thymic lymphoma between 1975 and 2018 from the nine states of the US were identified, including Atlanta, Connecticut, Detroit, Hawaii, Iowa, New Mexico, San Francisco-Oakland, Seattle–Puget Sound, and Utah. Incidence and mortality rates were analyzed using SEER*Stat 8.3.9 software. Univariate and multivariate Cox regressions were performed to identify prognostic factors. The Kaplan–Meier curve and log-rank test were used to compare overall survival (OS) among different treatments.ResultsA total of 233 patients diagnosed as thymic lymphoma were identified, and eight of them were lost to follow-up or died upon diagnosis. The incidence of thymic lymphoma was 2.032 per ten million (95% CI: 1.777–2.312), and the mortality rate was 0.649 per ten million (95% CI: 0.508–0.817). Among the 225 patients with definite follow-up, 98 were males and 127 were females, with a median age of 33 years. The Cox regression results showed that age and pathological type were independent risk prognostic factors. The 5-, 10-, and 20-year OS were 80.0%, 77.5%, and 70.9%, respectively. For Ann Arbor stage I and II patients, there was no significant difference between the surgical group (N = 78) and the non-operative group (N = 65; P = 0.270). The radiotherapy group (N = 79) had better OS than the non-radiotherapy group (N = 64) in the first 25 years, and the prognosis in the later years was not significantly different (P = 0.051). The chemotherapy group (N = 37) had a significantly better prognosis than the non-chemotherapy group (N = 37; P = 0.020). Patients who received postoperative radiotherapy (N = 45) or who only received radiotherapy (N = 34) seemed to have better OS than that of patients who only received surgery (N = 33), although the difference was not significant (P = 0.063).ConclusionsAge and pathological type were independent prognostic factors for thymic lymphoma. Surgical treatment had limited effects on OS, while both radiotherapy and chemotherapy could significantly improve the survival outcome.

Quantitative texture analysis based on dynamic contrast enhanced MRI for differential diagnosis between primary thymic lymphoma from thymic carcinoma

To evaluate the value of texture analysis based on dynamic contrast enhanced MRI (DCE-MRI) in the differential diagnosis of thymic carcinoma and thymic lymphoma. Sixty-nine patients with pathologically confirmed (thymic carcinoma, n = 32; thymic lymphoma, n = 37) were enrolled in this retrospective study. Ktrans, Kep and Ve maps were automatically generated, and texture features were extracted, including mean, median, 5th/95th percentile, skewness, kurtosis, diff-variance, diff-entropy, contrast and entropy. The differences in parameters between the two groups were compared and the diagnostic efficacy was calculated. The Ktrans-related significant features yielded an area under the curve (AUC) of 0.769 (sensitivity 90.6%, specificity 51.4%) for the differentiation between thymic carcinoma and thymic lymphoma. The Kep-related significant features yielded an AUC of 0.780 (sensitivity 87.5%, specificity 62.2%). The Ve-related significant features yielded an AUC of 0.807 (sensitivity 75.0%, specificity 78.4%). The combination of DCE-MRI textural features yielded an AUC of 0.962 (sensitivity 93.8%, specificity 89.2%). Five parameters were screened out, including age, Ktrans-entropy, Kep-entropy, Ve-entropy, and Ve-P95. The combination of these five parameters yielded the best discrimination efficiency (AUC of 0.943, 93.7% sensitivity, 81.1% specificity). Texture analysis of DCE-MRI may be helpful to distinguish thymic carcinoma from thymic lymphoma.

Nucks1 synergizes with Trp53 to promote radiation lymphomagenesis in mice.

NUCKS1 is a 27 kD vertebrate-specific protein, with a role in the DNA damage response. Here, we show that after 4 Gy total-body X-irradiation, Trp53+/− Nucks1+/− mice more rapidly developed tumors, particularly thymic lymphoma (TL), than Trp53+/− mice. TLs in both cohorts showed loss of heterozygosity (LOH) of the Trp53+ allele in essentially all cases. In contrast, LOH of the Nucks1+ allele was rare. Nucks1 expression correlated well with Nucks1 gene dosage in normal thymi, but was increased in the majority of TLs from Trp53+/− Nucks1+/− mice, suggesting that elevated Nucks1 message may be associated with progression towards malignancy in vivo. Trp53+/− Nucks1+/− mice frequently succumbed to CD4- CD8- TLs harboring translocations involving Igh but not Tcra/d, indicating TLs in Trp53+/− Nucks1+/− mice mostly originated prior to the double positive stage and at earlier lineage than TLs in Trp53+/- mice. Monoclonal rearrangements at Tcrb were more prevalent in TLs from Trp53+/− Nucks1+/− mice, as was infiltration of primary TL cells to distant organs (liver, kidney and spleen). We propose that, in the context of Trp53 deficiency, wild type levels of Nucks1 are required to suppress radiation-induced TL, likely through the role of the NUCKS1 protein in the DNA damage response.

Primary Hodgkin's disease of the thymus

The thymus is a lymphoid organ located in the antero-superior mediastinum and composed of heterogeneous admixture of lymphoid and epithelial elements. Thymic tumors are dominated by thymoma; lymhomas are uncommon involving the gland secondary to adenopathies. Primary thymic lymphomas are exceptional. We report a 25-year-old man referred to us because of chest pain and irritating cough since 2 months. Physical examination was normal and no lymph nodes were palpable. Chest radiography showed an anterior mediastinal mass. Computed tomography (CT) examination of the chest revealed a homogeneous tissular mass occupying the anterior mediastinum in its upper and medium stage, extending to the infundibulum without lymph node enlargement. Thymectomy and wedge were performed via a median sternotomy. Pathologic diagnosis was nodular sclerosis Hodgkin's disease of the thymus. The patient received 6 cycles of ABVD (Adriamycin [doxorubicin], bleomycin, vinblastin and dacarbazine) and mediastinal radiotherapy. Currently, tow years after diagnosis, he is well and free of disease. Primary thymic Hodgkin's lymphoma is a rare entity which should be included in the differential diagnosis of anterior mediastinal mass especially in young patients.

Epigenetic silencing of E- and N-cadherins in the stroma of mouse thymic lymphomas

Aberrant expression of some tumour suppressor genes and oncogenes by thymocytes had been involved in the development of primary thymic lymphomas induced by gamma-irradiation, but genetic alterations affecting critical genes expressed by stromal cells have not been yet explored. This paper analyzes a series of such tumours induced in C57BL/6J and in F1 hybrids of BALB/c and C57BL/6J mouse strains. As expected, hystopathological analyses revealed profound disorganizations within the thymus with a poor demarcation of the cortical and medullar areas. Immunological and quantitative on-line RT-PCR analyses confirm that E-cadherin (Cdh1) is essentially expressed by stromal cells of the thymus, while evidencing that the expression of this gene is significantly reduced in all tumours. In addition, and contrary to what one would expect, N-cadherin (Cdh2) that is exclusively expressed by stromal cells is likewise down-regulated in most of the thymic lymphomas. Although hypermethylation of the promoter region appears to be involved in the inactivation of Cdh2 in all tumours, additional epigenetic mechanisms mediated by repressors such as Snai1 may also play a role in Cdh1 silencing. These results represent the first reported case for tumour-associated gene alterations occurring not in the tumour cells per se, but in the stromal cells of primary thymic lymphomas. Additionally, since the expression of both genes is significantly up-regulated after a single high dose of gamma-radiation, but remained unchanged in treated thymic-lymphoma-free-mice, epigenetic down-regulation of E- and N-cadherin appears to occur concomitantly with the progression towards the most advanced stages of gamma-radiation-induced thymic lymphomas.

Evidence of a possible epigenetic inactivation mechanism operating on a region of mouse chromosome 19 in gamma-radiation-induced thymic lymphomas.

Loss of heterozygosity (LOH) analysis, performed in 68 gamma-radiation-induced primary thymic lymphomas of F1 hybrid mice, provided evidence of significant LOH on chromosome 19 in a region defined by the D19Mit106 (22 cM) and D19Mit100 (27 cM) markers (Thymic Lymphoma Suppressor Region 8, TLSR8). Cd95 and Pten, two genes mapped at this region, were inactivated in a vast majority of these tumors (85.3% for Cd95 and 61.8% for Pten). Moreover, altered expression of Cd95 and Pten occurred concomitantly in 34 of 68 (50%) thymic lymphomas suggesting a coordinated mechanism of inactivation of these genes. Surprisingly, we also found that Jak2, a proto-oncogene located between Cd95 and Pten, was simultaneously inactivated in a significant fraction of the tumors analysed (24 of 34, 70.6%). Taken together these findings and the lack of mutations in the coding sequences of the mentioned genes clearly suggest a possible regional epigenetic inactivation mechanism on mouse chromosome 19 operating during the development of these tumors.

Thymic lymphomas induced by N-propyl-N-nitrosourea (PNU) in the BUF/Mna rat, an inbred strain with a high incidence of spontaneous thymoma.

N-Propyl-N-nitrosourea (PNU) is known to be a strong leukemogen, inducing myelogenous leukemia or thymic lymphoma in some strains of rat. The thymic lymphomagenic effect of PNU has been demonstrated in F344 rats. On the other hand, the BUF/Mna rat has been established as an inbred strain that develops spontaneous thymomas after one year of age. In the present experiment, PNU was continuously administered in drinking water to male and female BUF/Mna rats starting at 5 weeks of age. Thymic lymphomas were induced in all PNU-treated rats with an average latent period as short as 14 experimental weeks. These results show the high susceptibility of the BUF/Mna rat to the lymphomagenic activity of PNU. The BUF/Mna rat is an ideal strain for studies on epithelial cell-lymphocyte interaction, not only in the development of thymic lymphomas but also in that of spontaneous thymoma. Karyotypes of twelve primary thymic lymphomas induced by PNU were analyzed for chromosomal abnormalities. Chromosomal abnormalities were often found in chromosomes 11 and 2. In some types of abnormality, dup (11q) and del(2q) were most frequently observed. In addition, trisomy of chromosome 7, on which the c-myc gene is mapped, was observed in five lymphomas, and monosomy of chromosomes 20 and X in six and five cases, respectively, though these changes were generally observed in a minor cell population in each case.

Characterization of cell markers in type B retroviral-induced thymic lymphomas—II. Surface antigen phenotype, karyotype and proviral integration pattern in cultured lymphoma cells and cloned lines

This study examined the surface antigen phenotype, karyotype and proviral integration patterns of cultured cells from murine thymic lymphomas induced by injecting neonatal mice with the type B leukemogenic retrovirus (DMBA-LV). Cells from the primary thymic lymphomas were established in mass culture and from these, clonal tumor cell lines were derived. During in vitro culture of lymphoma cells, Lyt 1 −2 + cells predominated with an apparent selection against cells of the Lyt 1 +2 − and Lyt 1 +2 + phenotypes. Of 21 cloned lines established, five had a diploid chromosome complement and expressed the Lyt 1 −2 + phenotype. The other 16 clones lines were characterized by trisomy of chromosome 15 and expressed the Lyt 1 +2 + or Lyt 1 −2 + phenotype. Cells characterized by either a diploid or trisomy chromosome complement were capable of growth in vivo. Southern blot analyses showed that during growth in culture, cells from the mass cultures and cloned lines continued to acquire low numbers of new integrated DMBA-LV proviral copies while maintaining the basic proviral integration pattern present in the DNA from cells of the primary lymphomas. These findings support the notion that the acquisition of new genetic information in cells from DMBA-LV-induced thymic lymphomas may contribute to the continual generation of tumor heterogeneity.

Integration of type B retroviral DNA in virus-induced primary murine thymic lymphomas.

In previous studies we described the isolation and characterization of a highly leukemogenic virus, DMBA-LV, isolated from a transplanted, chemical carcinogen-induced thymic lymphoma. The virus is composed of a mixture of two unrelated retroviral genomes, one highly related to type B milk-borne mouse mammary tumor virus isolates and the other partially related to type C viral genomes. In the present study, primary thymic lymphomas induced by DMBA-LV in CFW/D, NIH Swiss, C3H/Bi/Ka, and C57BL/Ka mice were assessed for the presence of newly integrated type B retroviral DNA. All 46 primary thymic lymphomas examined contained one to four newly acquired murine mammary tumor virus proviruses. Based on the sizes of provirus-cell DNA junction fragments, the integration of newly acquired murine mammary tumor virus proviruses did not appear to be random.

Chromosomal alterations in the course of viral leukemogenesis in the rat

A cytogenetic study of 35 primary thymic lymphomas induced in rats by the Gross leukemia virus revealed the presence of widespread aneuploidies, ranging from cells with 35 chromosomes or less to triploid and tetraploid cells. Analysis by G banding revealed that chromosomes No. 1, 2, 9, 12, 18–20, X in females, and Y in males were lost preferentially; unidentified marker chromosomes of various morphology and length were present in a high percentage of aneuploid mitoses. Similar alterations were observed in preleukemic thymi from rats sacrificed 30 days after virus injection, when clinical or histologic signs of the disease had not yet appeared. In rats that were sacrificed 7 or 15 days after virus injection, chromosomal anomalies were much more limited, although significantly more abundant than in the controls. Rats that were free of tumor at 130–160 days after virus injection showed a regression of the chromosomal anomalies, suggesting that these are not irreversible and are not necessarily followed by the development of a lymphoma.

The Ratio of Cytochromes in Radiation-Induced Leukemia Cells of Mice

The content of cytochromes in various leukemia cells and normal thymocytes of mice was determined spectrophotometrically at room temperature and at the temperature of liquid nitrogen. The ratio of cytochrome c + c/sub 1/ to cytochrome aa/sub 3/ of primary thymic lymphomas, whether arising spontaneously in AKR mice or induced by x-irradiation in NIH Swiss mice, was similar to that of normal thymocytes. In contrast, three lines of transplantable ascites lymphoma cells, originally derived from radiation-induced lymphoma, showed a definitely higher ratio than that of normal thymocytes. The results revealed that the c + c/sub 1//aa/sub 3/ ratio of the lymphoma cells increased in successive generations during serial passage of the cells in vivo.