Perineural invasion on biopsy is associated with upstaging at radical prostatectomy in Gleason score 3 + 4 = 7 prostate cancer

This study assesses if perineural invasion (PNI) detected on biopsy with Gleason score (GS) 3 + 4 = 7 prostate cancer (PCa) is associated with upstaging/upgrading of disease after radical prostatectomy (RP). 154 patients with GS 3 + 4 = 7 PCa diagnosed from biopsy who underwent RP were assessed for PNI. The percentage of biopsy sites with PNI (%PNI) was also calculated. Pattern 4 morphologies (ill‐defined glands [IDG], fused, cribriform, and glomerulations) were also assessed. Clinical information, GS and stage after RP were retrieved from the medical records. 45 % (69/154) of patients were upstaged (≥pT3) and 29 % (44/154) were upgraded to GS >3 + 4 = 7 after RP. 37 % (57/154) of patients had PNI which was associated with upstaging (RR 1.4; P = 0.04) but not upgrading (RR 0.9; P = 0.7). There was higher %PNI in upstaged patients (12.1 % ± 1.8 vs. 7.1 % ± 1.5, P = 0.03) with a significant correlation between %PNI and ≥pT3 (r = 0.178, P = 0.027). After multivariate analysis, only cribriform formations were significantly associated with upstaging (P = 0.009). The presence of PNI in biopsies with GS 3 + 4 = 7 PCa is associated with upstaging at RP but is a weaker predictor of ≥pT3 disease than cribriform morphology.     

Migrant μ+δ+ and static μ+δ− B lymphocyte subsets

Immunoglobulin isotype expression in isolated lymph node (LN), spleen and blood lymphocyte suspensions was assessed in rats. The proportion of μ⁺δ^? B cells in spleen (34%) was approximately twice that in blood and LN. Immunohistological examination of spleens showed the cells of the marginal zones to be predominantly μ⁺δ^?. On the other hand, μ⁺δ⁺ B cells were mainly confined to the follicles in both spleen and LN. These follicles had a minor μ⁺δ^? component. The migratory properties of B cells with these two phenotypes were assessed by depleting lymphocytes migrating through the white pulp of rat spleen. This was achieved by placing a ³²P-impregnated β-emitting polythene strip over one half of the spleen. Examination of the nonirradiated half of the spleen, LN and peripheral blood after 12 days irradiation showed selective loss of δ⁺ B cells. The μ⁺δ^? cells of the splenic marginal zone were numerically unaltered. There was also a substantial residual μ⁺δ^? B cell presence in the small lymphocyte compartment of follicles of LN and spleen in depleted animals. In addition, the blood selectively retained a μ⁺δ^? B cell component. This was not derived from the spleen, as μ⁺δ^? blood B cell numbers were sustained even where both halves of the spleen were irradiated. It is concluded that: (a) the major static B cell component of spleen and LN is μ⁺δ^?, (b) that most if not all δ⁺ B cells repeatedly migrate through the spleen and (c) there is a blood-born μ⁺δ^? component which is resistant to depletion by splenic irradiation.     

Outcomes of Gleason score 3 + 4 = 7 prostate cancer with minimal amounts (<6%) vs ≥6% of Gleason pattern 4 tissue in needle biopsy specimens

ObjectiveThe International Society of Urological Pathology Gleason grading system was modified in 2005. Since the modified system was introduced, many cancers that previously would have been categorized as Gleason score (GS) 6 are now categorized as GS 7 based on biopsy specimens that only contain minimal amounts (<6%) of Gleason pattern (GP) 4 tissue. However, the clinical significance of observing <6% of GP 4 tissue in biopsies of GS 7 prostate cancer has not been studied.Material and methodsThis study was based on needle biopsy specimens that were categorized as GS 6 or GS 7 and were obtained from patients who underwent radical prostatectomy (RP) with available follow-up data. We assessed the quantity of GP 4 tissue in biopsy specimens of GS 7 prostate cancer. Further, we evaluated the correlation between the quantity of GP 4 tissue and disease progression after RP.ResultsGP 4 comprising 26–49% of the specimen, GS 4+3 and percentage of total core tissue scored as positive were significant and independent predictors of prostate-specific antigen (PSA) failure after RP, as assessed using a multivariate Cox regression model that included the quantity of GP 4 in the prostate biopsy specimen, preoperative PSA, perineural invasion, clinical stage, number of positive cores, and percentage of core tissue scored as positive. Cases with GS 3+3 and cases in which the observed GP 4 area was <6% did not differ significantly in terms of biochemical PSA recurrence (BPR) status. In contrast, cases with 6–25% GP 4 tissue, 26–49% GP 4 tissue, and GS 4+3 showed more frequent BPR than cases with GS 3+3.ConclusionsOur data suggest that the quantity of GP 4 tissue in GS 7 cancer has clinical significance. However, there is a need for larger studies of the clinical significance of biopsy specimens that include <6% GP 4 tissue. We should reconsider whether the amount of GP 4 should be included in standart pathology reports.     

Comparative influence of cribriform growth and percent Gleason 4 in prostatic biopsies with Gleason 3+4 cancer

The International Society of Urological Pathology endorses specifying presence of cribriform architecture in Gleason (G)4 prostate cancer because of cribriform's aggressiveness. The relative effect of cribriform presence versus percentage G4 within grade group (GG)2 or 3 was uncertain.194 men's biopsies with GG2 with or without cribriform (excluding glomeruloid from cribriform) and GG3 without cribriform (controls) from 4 years were reviewed. 173 cases had follow-up including 147 GG2 (15/147 or 10% had cribriform) and 26 GG3. Effects of total tumor specimen involvement, %Gleason 4, and cribriform were stratified into prostatectomy (n = 90), radiotherapy (n = 61), and watching waiting (n = 22) groups.Median follow-up duration was 3.32 years (range 1.90–6.18). Biochemical failures in the above 3 cohorts numbered 9 (9/90; 10%), 5 (5/61; 8%), and 13 (13/22; 59%) respectively. In all groups, (GG2+ GG3, n = 173), the HR for C pattern was 1.64. In GG2, cribriform presence (considering glomeruloid as non-cribriform) conferred a hazard ratio (HR) of 1.51 (p = 0.48). It was 1.38, excluding glomeruloid. In watchful waiting cohort only, presence of C conferred a HR of 2.62 (p = 0.086). All remaining comparisons including percent G4, remained not significant.Thus, only in WW group did cribriform pattern presence approach significance. Detection of differences otherwise was not feasible, probably because: 1) biochemical failure is too rare in GG2 cancer; 2) cribriform frequency was only 10% in GG2 (in current study), less than in higher-grade cancer. 3) Use of biopsy tissue is subject to sampling variation which may undersample cribriform pattern, though biopsy forms the basis of treatment decisions.     

Aneusomy of chromosomes 7, 8, and 17 and amplification of HER-2/neu and epidermal growth factor receptor in Gleason score 7 prostate carcinoma: a differential fluorescent in situ hybridization study of Gleason pattern 3 and 4 using tissue microarray.

Recent evidence shows that the proportion of poorly differentiated prostate carcinoma (Gleason pattern [GP] 4/5) is a surrogate factor for biochemical failure after radical prostatectomy (RP). However, little is known about specific molecular and cytogenetic changes in this aggressive component of localized prostate cancer. We constructed a tissue microarray containing areas of GP 3 and 4 from formalin-fixed radical prostatectomy specimens of 39 patients with Gleason score 7 carcinoma (>or=50% GP 4), known pathologic staging parameters (stage < T3b), and biochemical failure data (mean follow-up, 30 months; range, 5 to 74 months). Interphase fluorescent in situ hybridization (FISH) was performed on 5-microm microarray sections using pericentromeric probes to chromosomes 7, 8, and 17 and probes for the HER-2/neu and epidermal growth factor receptor (EGFR) genes. Low-level amplification of HER-2/neu was found in 26% of cases (3 to 5 signals per nucleus, corrected for chromosome 17 aneusomy). Aneusomy of chromosomes 7, 8, and 17 was identified in 21%, 15%, and 5% of cases, respectively. All aberrations occurred almost exclusively in GP 4 carcinoma (8 of 8 aneusomies 7, 2 of 2 trisomies 17, 9 of 10 HER-2/neu amplifications, and 5 of 6 aneusomies 8; P < .001). The presence of HER-2/neu amplification was associated with high tumor volume (>2.0 cm(3), P = 0.004). Among patients with negative surgical margins, gain of chromosome 7 was associated with biochemical failure after RP (P =.004, log-rank). Amplification of the EGFR gene occurred in only 1 case (3%). Significant differences in HER-2/neu amplification and gain of chromosomes 7, 8, and 17 were detected between GP 4 prostate carcinoma and GP 3. The frequency of aberrations increased with tumor volume. Chromosome 7 abnormalities may play an important role in cancer progression in margin-negative patients. EGFR amplification was rare, suggesting that this oncogene is not altered at the gene copy number level.     

CD30 + T cell enriched primary cutaneous CD4 + small/medium sized pleomorphic T cell lymphoma: A distinct variant of indolent CD4 + T cell lymphoproliferative disease

Primary cutaneous CD4 + small/medium sized pleomorphic T-cell lymphoma (SMPTCL) is unique among the peripheral T-cell lymphomas because of its indolent nature, typically presenting as a solitary nodule or plaque in the head and neck area of middle-aged and older adults. Recent studies have suggested a follicular helper cell origin for these lesions.Materials and methodsA retrospective review was conducted on all cases of SMPTCL diagnosed between 2008 and 2017. The goal of our study was to better categorize the clinical, pathologic and molecular features of cases of SMPTCL showing a significant degree of CD30 neoplastic large cell infiltration.ResultsFifteen patients (10 male, 5 female) were encountered (age 33–86 years at presentation). All lesions were solitary and the head and neck region was the most common area of involvement (7 cases). Surgical excision alone was performed in 6 cases and was supplemented with radiation in 5 cases. Disease recurrence did not occur. Spontaneous regression following biopsy was reported and two patients had a history compatible with lymphomatoid papulosis. All cases showed pathologic features characteristic of SMPTCL. Additionally, there were many larger CD30 + T-cells occupying 15–30% of the infiltrate. Monoclonality was demonstrated in 5 of 10 cases in which clonality studies were performed.ConclusionCD30 positivity amidst large neoplastic T-cells is not uncommon in SMPTCL. The extent of CD30 positivity in SMPTCL needs to be defined further along with its association with other forms of CD30 + lymphoproliferative disease including its potential categorization as a form of endogenous CD30 + lymphoproliferative disease.     

Aneusomy of chromosomes 7, 8, and 17 and amplification of HER-2/neu and epidermal growth factor receptor in Gleason score 7 prostate carcinoma: A differential fluorescent in situ hybridization study of Gleason pattern 3 and 4 using tissue microarray

Recent evidence shows that the proportion of poorly differentiated prostate carcinoma (Gleason pattern [GP] 4/5) is a surrogate factor for biochemical failure after radical prostatectomy (RP). However, little is known about specific molecular and cytogenetic changes in this aggressive component of localized prostate cancer. We constructed a tissue microarray containing areas of GP 3 and 4 from formalin-fixed radical prostatectomy specimens of 39 patients with Gleason score 7 carcinoma (≥50% GP 4), known pathologic staging parameters (stage < T3b), and biochemical failure data (mean follow-up, 30 months; range, 5 to 74 months). Interphase fluorescent in situ hybridization (FISH) was performed on 5-μm microarray sections using pericentromeric probes to chromosomes 7, 8, and 17 and probes for the HER-2/neu and epidermal growth factor receptor (EGFR) genes. Low-level amplification of HER-2/neu was found in 26% of cases (3 to 5 signals per nucleus, corrected for chromosome 17 aneusomy). Aneusomy of chromosomes 7, 8, and 17 was identified in 21%, 15%, and 5% of cases, respectively. All aberrations occurred almost exclusively in GP 4 carcinoma (8 of 8 aneusomies 7, 2 of 2 trisomies 17, 9 of 10 HER-2/neu amplifications, and 5 of 6 aneusomies 8; P < .001). The presence of HER-2/neu amplification was associated with high tumor volume (>2.0 cm³, P = 0.004). Among patients with negative surgical margins, gain of chromosome 7 was associated with biochemical failure after RP (P = .004, log-rank). Amplification of the EGFR gene occurred in only 1 case (3%). Significant differences in HER-2/neu amplification and gain of chromosomes 7, 8, and 17 were detected between GP 4 prostate carcinoma and GP 3. The frequency of aberrations increased with tumor volume. Chromosome 7 abnormalities may play an important role in cancer progression in margin-negative patients. EGFR amplification was rare, suggesting that this oncogene is not altered at the gene copy number level. H P 32:1392-1397. Copyright © 2001 by W.B. Saunders Company     

Imaging of nuclear Overhauser enhancement at 7 and 3 T

Nuclear Overhauser enhancement (NOE) is a type of magnetization transfer using cross‐relaxation. It originates from mobile macromolecules, which may have relevance to the evaluation of tumor features. We studied the value of NOE imaging at 7 and 3 T and suggest a utility for diagnosing human brain tumors. Two types of protein solution at different concentrations and pH values, and six normal Sprague Dawley (SD) rats, were used to detect NOE signal with a 7 T scanner. Then, six healthy volunteers and 11 patients with brain tumors (six gliomas and five meningiomas) were included at 3 T. Z‐spectra were measured and NOE weighted (NOE*) images were acquired with a three‐offset measurement. Wide spectral separation was shown at both 7 T and 3 T delineating the NOE peak in the Z‐spectrum. The concentration dependence and pH independence of NOE were confirmed in phantom experiments, and NOE values were greater in white matter than in gray matter in vivo. At 3 T, data indicated that NOE* maps were slightly hypointense in gliomas and were not obviously different from meningiomas. Thus, NOE imaging may help distinguish benign from malignant tumors, and as such may contribute to diagnosing brain tumors.     

前列腺癌8号染色体改变与Gleason评分之间的相关性

目的探索前列腺癌8号染色体数目增加及c-myc基因和脂蛋白脂酶（LPL）基因状态和发生频率,分析8号染色体改变与前列腺癌Gleason评分之间的相关性及其在前列腺癌发生发展中的作用。方法采用ProVysion^TM三色探针组合,以荧光原位杂交（FISH）方法检测34例未经临床治疗的前列腺癌穿刺组织石蜡切片标本的8号染色体改变,其中包括Gleason评分5分者1例,6分者10例,7分者14例,8分者4例,9分者5例,并进行8号染色体各种异常之间及其与前列腺癌Gleason评分级别之间的关联性分析。结果8号染色体增加为17／34（50％）,c-myc基因拷贝数增加为21／34（61．8％）,LPL单体为15／34（44．1％）,c-myc基因扩增为23／34（67．6％）,LPL基因缺失为21／34（61．8％）,同时具有LPL基因缺失和c-myc基因扩增为16／34（47．1％）,至少有其中一种遗传学异常者为29／34（85．3％）。8号染色体增加与Gleason评分级别增高呈明显的正相关关系（P=0．0006）;c-myc基因拷贝数增加与Gleason评分级别增高呈正相关关系（P=0．0035）;LPL缺失与Gleason评分级别增高呈负相关关系（P=0．0383）;调整年龄后,除了上述三个变量与Gleason评分级别的相关关系仍然存在以外,c-myc基因扩增与Gleason评分级别增高也呈现正相关关系（P=0．0462）。结论8号染色体数目增加、c-myc基因拷贝数增加、c-myc基因扩增和LPL基因丢失都与Gleason评分级别有关,c-myc基因扩增同时伴有LPL基因缺失也是前列腺癌的遗传学特征之一,提示8号染色体异常可能与前列腺癌的发生和进展有关。     

不同标本前列腺癌Gleason评分差异分析

目的：探讨不同标本前列腺癌Gleason评分（Gleason score,GS）的差异。方法：研究前列腺癌GS系统主要指标均值及所占比例,比较穿刺活检（needle biopsy,NB）、经尿道前列腺切除（transurethral resection of the prostate,TURP）以及前列腺切除（prostatectomy,PC）标本中的差异。结果：NB,TURP和PC中GS均值分别为7.14±1.11,7.14±1.32和6.78±0.91,PC中GS均值显著低于NB（P=0.001）和TURP（P=0.029）。GS 5,6,7,8,9和10分在NB中分别占0.4%,33.9%,34.7%,15.3%,13.4%和2.3%;在TURP中分别占4.3%,40.0%,18.6%,17.1%,14.3%和5.7%;在PC中分别占2.6%,39.1%,40.4%,11.3%,6.6%和0;GS 5分在TURP中所占比例显著高于NB（P=0.001）,在PC中显著高于NB（P=0.014）;GS 8分在TURP中所占比例显著高于NB（P=0.007）和PC（P=0.001）;GS 9分在TURP中所占比例显著高于PC（P=0.025）;GS 10分在TURP中所占比例显著高于PC（P=0.003）。结论：不同标本前列腺癌GS存在差异;在三种标本中PC GS最低,TURP GS偏高;前列腺癌早期发现和早期治疗还需要进一步加强。     

Low Gleason score prostatic adenocarcinomas are no longer viable entities

Gleason scoring is the most powerful predictor of the behaviour of prostatic adenocarcinoma. It was devised more than 30 years ago and is used by pathologists worldwide. However, advances in immunochemistry and the understanding of the biological potential of some tumours have made the diagnosis of tumours of grades 1 and 2, and hence low score (2-4), a rarity. It is suggested that Gleason score 2(1 + 1) tumours, if they do exist, are so rare and so benign that they are a redundant entity and should not be identified as a malignancy. Similar problems revolve around the diagnosis of Gleason score 4(2 + 2) tumours and the criteria for their diagnosis are difficult to evaluate and revolve around reference to the score 2(1 + 1) lesion. The continued presence of low-grade tumours in the Gleason grading system leads to diagnostic confusion, potential error and, more importantly, confusion for patients diagnosed with score 6(3 + 3) tumours. I suggest that Gleason grade 1 (and therefore scores of 2 and 3) should be removed from the Gleason grading system. Gleason grade 2 (and therefore score 4 and 5) tumours require serious re-evaluation if they are to remain clinically useful diagnoses.     

Interobserver reproducibility of modified Gleason score in radical prostatectomy specimens

The Gleason score (GS) of prostate cancer is calculated by adding primary and secondary Gleason grades with patterns occupying less than 5% of the tumour often not included despite their probable prognostic significance. A modified Gleason score (mGS) comprising primary and tertiary patterns of higher grade has been proposed, but its interobserver variability has yet to be elucidated. Slides from 69 consecutive prostatectomy specimens were circulated among four genitourinary pathologists. GS and mGS were assessed, and results were compared in pairs. Mean weighted kappa for GS and mGS were 0.56 (range 0.52–0.66) and 0.58 (range 0.49–0.74), respectively. The difference between GS and mGS was 0, 1 and 2 score units in 66%, 26% and 8%, respectively, mean 0.41 score units (range 0.24–0.51). The increment was greater for transition-zone tumours than for peripheral-zone tumours (0.63 and 0.35 score units, respectively, P=0.002). An odd mGS (5, 7 or 9) was more often given than an odd GS (77% and 62%, respectively, P<0.001). Disagreement between observers greater than 1 score unit was more common with mGS than GS (18% and 4%, respectively, P<0.001). In conclusion, overall mean weighted kappa for interobserver reproducibility of mGS is at least as high as that of GS. However, there is a clustering of mGS in odd scores, and severe disagreement is more commonly observed than with GS. Training of mGS assessment would possibly improve agreement. Tertiary Gleason patterns need to be better defined.The study was approved by the ethics committee of The Karolinska Institute (01–443).     

CD4<Superscript>+</Superscript>FOXP3<Superscript>+</Superscript> T regulatory cells decrease and CD3<Superscript>+</Superscript>CD8<Superscript>+</Superscript> T cells recruitment in TILs from melanoma metastases after electrochemotherapy

Electrochemotherapy (ECT) represents an effective local treatment for skin unresectable melanoma metastases with high overall objective response rate. ECT is based on the combination of anti-neoplastic drugs administration and cancer cells electroporation. Whether ECT can also activate the immune system is a matter of debate, however a significant recruitment of dendritic cells in melanoma treated metastases has been described. Herein we investigated immediate and late effects of ECT treatment on T cell subsets in ECT-treated lesions by fluorescent immunohistochemistry. Biopsies from melanoma patients (n = 10) were taken before ECT (t0), at d1 and d14 from treatment. At t0, CD3⁺CD4⁺ T cells were the most represented T cells, well detected in the perilesional dermis, particularly at tumour margin, while CD3⁺CD8⁺ T cells were less represented. CD4⁺FOXP3⁺ T regulatory (Treg) cells were present in the perilesional dermis and within the lesion. ECT induced a significant decrease of CD4⁺FOXP3⁺ Treg cells percentage in the perilesional dermis, observed at d1 and at d14 (p < 0.001). CD3⁺CD8⁺ T cells frequency significantly increased at d14 from treatment in the perilesional dermis (p < 0.001). Furthermore calreticulin translocation to the plasma membrane, a hallmark of immunogenic cell death, was observed in metastatic cells after ECT. The data reported here confirm that ECT induces a local response, with a lymphoid infiltrate characterized by CD4⁺FOXP3⁺ Treg cells decrease and CD3⁺CD8⁺ T cells recruitment in the treated lesions. These results might contribute to design novel combinational therapeutic approaches with ECT and immunotherapy in order to generate a systemic long-lasting anti-melanoma immunity.     

A study of Gleason score interpretation in different groups of UK pathologists; techniques for improving reproducibility

AIMS: To test the effectiveness of a teaching resource (a decision tree with diagnostic criteria based on published literature) in improving the proficiency of Gleason grading of prostatic cancer by general pathologists. METHODS: A decision tree with diagnostic criteria was developed by a panel of urological pathologists during a reproducibility study. Twenty-four general histopathologists tested this teaching resource. Twenty slides were selected to include a range of Gleason score groups 2-4, 5-6, 7 and 8-10. Interobserver agreement was studied before and after a presentation of the decision tree and criteria. The results were compared with those of the panel of urological pathologists. RESULTS: Before the teaching session, 83% of readings agreed within +/- 1 of the panel's consensus scores. Interobserver agreement was low (kappa = 0.33) compared with that for the panel (kappa = 0.62). After the presentation, 90% of readings agreed within +/- 1 of the panel's consensus scores and interobserver agreement amongst the pathologists increased to kappa = 0.41. Most improvement in agreement was seen for the Gleason score group 5-6. CONCLUSIONS: The lower level of agreement among general pathologists highlights the need to improve observer reproducibility. Improvement associated with a single training session is likely to be limited. Additional strategies include external quality assurance and second opinion within cancer networks.