Fumitake Hata

Phase I clinical study of anti-apoptosis protein, survivin-derived peptide vaccine therapy for patients with advanced or recurrent colorectal cancer.

Survivin is a member of the inhibitor of apoptosis protein (IAP) family containing a single baculovirus IAP repeat domain. It is expressed during fetal development but becomes undetectable in terminally differentiated normal adult tissues. We previously reported that survivin and its splicing variant survivin-2B was expressed abundantly in various types of tumor tissues as well as tumor cell lines and was suitable as a target antigen for active-specific anti-cancer immunization. Subsequently, we identified an HLA-A24-restricted antigenic peptide, survivin-2B80-88 (AYACNTSTL) recognized by CD8+ cytotoxic T lymphocytes (CTLs). We, therefore, started a phase I clinical study assessing the efficacy of survivin-2B peptide vaccination in patients with advanced or recurrent colorectal cancer expressing survivin. Vaccinations with survivin-2B peptide were given subcutaneously six times at 14-day intervals. Of 15 patients who finished receiving the vaccination schedule, three suffered slight toxicities, including anemia (grade 2), general malaise (grade 1), and fever (grade 1). No severe adverse events were observed in any patient. In 6 patients, tumor marker levels (CEA and CA19-9) decreased transiently during the period of vaccination. Slight reduction of the tumor volume was observed in one patient, which was considered a minor responder. No changes were noted in three patients while the remaining eleven patients experienced tumor progression. Analysis of peripheral blood lymphocytes of one patient using HLA-A24/peptide tetramers revealed an increase in peptide-specific CTL frequency from 0.09% to 0.35% of CD8+ T cells after 4 vaccinations. This phase I clinical study indicates that survivin-2B peptide-based vaccination is safe and should be further considered for potential immune and clinical efficacy in HLA-A24-expression patients with colorectal cancer.

A Potent Immunogenic General Cancer Vaccine That Targets Survivin, an Inhibitor of Apoptosis Proteins

We reported previously a HLA-A24-restricted antigenic peptide, survivin-2B80-88 (AYACNTSTL), recognized by CD8+ CTL. This peptide was derived from survivin protein, an inhibitor of apoptosis proteins, expressed in a variety of tumors, such as adenocarcinoma, squamous cell carcinoma, and malignant melanoma. In this report, we provide further evidence that survivin-2B80-88 peptide might serve as a potent immunogenic cancer vaccine for various cancer patients. Overexpression of survivin was detected in surgically resected primary tumor specimens of most breast and colorectal cancers and some gastric cancers as assessed by immunohistochemical study. HLA-A24/survivin-2B80-88 tetramer analysis revealed that there existed an increased number of CTL precursors in peripheral blood mononuclear cells (PBMC) of HLA-A24+ cancer patients, and in vitro stimulation of PBMCs from six breast cancer patients with survivin-2B80-88 peptide could lead to increases of the CTL precursor frequency. Furthermore, CTLs specific for this peptide were successfully induced from PBMCs in all 7 (100%) patients with breast cancers, 6 of 7 (83%) patients with colorectal cancers, and 4 of 7 (57%) patients with gastric cancers. These data indicate that survivin expressed in tumor tissues is antigenic in cancer patients, and survivin-2B80-88-specific CTLs are present in PBMCs of various cancer patients. Our study raises the possibility that this peptide may be applicable as a general cancer vaccine to a large proportion of HLA-A24+ cancer patients.

Clinical and immunological evaluation of anti-apoptosis protein, survivin-derived peptide vaccine in phase I clinical study for patients with advanced or recurrent breast cancer

BackgroundWe previously reported that survivin-2B, a splicing variant of survivin, was expressed in various types of tumors and that survivin-2B peptide might serve as a potent immunogenic cancer vaccine. The objective of this study was to examine the toxicity of and to c linically and immunologically evaluate survivin-2B peptide in a phase I clinical study for patients with advanced or recurrent breast cancer.MethodsWe set up two protocols. In the first protocol, 10 patients were vaccinated with escalating doses (0.1–1.0 mg) of survivin-2B peptide alone 4 times every 2 weeks. In the second protocol, 4 patients were vaccinated with the peptide at a dose of 1.0 mg mixed with IFA 4 times every 2 weeks.ResultsIn the first protocol, no adverse events were observed during or after vaccination. In the second protocol, two patients had induration at the injection site. One patient had general malaise (grade 1), and another had general malaise (grade 1) and fever (grade 1). Peptide vaccination was well tolerated in all patients. In the first protocol, tumor marker levels increased in 8 patients, slightly decreased in 1 patient and were within the normal range during this clinical trial in 1 patient. With regard to tumor size, two patients were considered to have stable disease (SD). Immunologically, in 3 of the 10 patients (30%), an increase of the peptide-specific CTL frequency was detected. In the second protocol, an increase of the peptide-specific CTL frequency was detected in all 4 patients (100%), although there were no significant beneficial clinical responses. ELISPOT assay showed peptide-specific IFN-γ responses in 2 patients in whom the peptide-specific CTL frequency in tetramer staining also was increased in both protocols.ConclusionThis phase I clinical study revealed that survivin-2B peptide vaccination was well tolerated. The vaccination with survivin-2B peptide mixed with IFA increased the frequency of peptide-specific CTL more effectively than vaccination with the peptide alone, although neither vaccination could induce efficient clinical responses. Considering the above, the addition of another effectual adjuvant such as a cytokine, heat shock protein, etc. to the vaccination with survivin-2B peptide mixed with IFA might induce improved immunological and clinical responses.

Septic Thrombophlebitis of the Portal and Superior Mesenteric Veins as a Complication of Appendicitis: Report of a Case

Pylephlebitis is extremely rare and associated with high mortality, even in this modern era. It usually occurs secondary to infection in the region drained by the portal systems or in the structure contiguous to the portal vein. We report a case of septic thrombophlebitis of the portal and superior mesenteric veins (SMV) with multiple liver abscesses caused by acute appendicitis with an abscess of the mesoappendix. We performed appendectomy and successfully removed the thrombi using a Fogarty catheter. Postoperative histopathological examination confirmed a diagnosis of appendicitis and septic thrombophlebitis of the portal vein and SMV. The patient recovered completely with appropriate medical and surgical treatment.

Identification of segments VI and VII of the liver based on the ramification patterns of the intrahepatic portal and hepatic veins.

We describe the pattern of intrahepatic vessel ramification in the right posterior hepatic sector in a population of 197 adults. Each specimen was dissected from its visceral (inferior) surface in order to demonstrate variations in the distribution of the portal vein branches to the hepatic segments of the right lobe, especially to segments VI (S6) and VII (S7) as described by Couinaud. We also examine whether three hepatic veins, i.e., the right hepatic vein (RHV), middle hepatic vein (MHV), and the short hepatic vein (SHV), aid the identification of segmental portal branches in the lower posterior sector. Four major patterns of branching of the posterior sectorial trunk of the portal vein system are described. In group A (32.0%) a single posterior trunk formed an arch‐like pattern sending multiple branches to S6 and S7 (P6 and P7). We named the multiple branches to the apparent S6 the inferoposterior portal branches. It was difficult to identify which of these branches were equivalent to P6. In group B (27.9%), the posterior sectorial trunk bifurcated to form P6 and P7. In most of the specimens in this group, therefore, we were able clearly to identify both S6 and S7 based on the portal vein system. In group C (6.6%), the trunk trifurcated to form P6, P7, and an intermediate branch, which supplied both segments or a gray zone between them. Group D (33.5%) included variations of the anterior segmental branches, and in specimens of this group, the anteromedial border of the sector was difficult to identify. Notably, the three‐dimensional interdigitating topographical relationship of the hepatic veins and the portal branches was not evident in the lower posterior sector, since tributaries of the RHV and the portal branches followed similar courses and paralleled each other in the region and since the territory of the SHV was usually restricted to the superficial parenchyma near the inferior surface. In group A, tributaries of the RHV/SHV (>3 mm in diameter) passed between the inferoposterior portal branches in only 22.2%/14.3% of the specimens. Thus the hepatic veins often did not reveal which of the multiple inferoposterior branches was P6. Moreover, in the subset of Group B in which the segments were identified based on the portal vein ramification, tributaries of the RHV/SHV (>3 mm in diameter) showed the intersegmental interdigitating arrangement in only 32.0%/6.0% of the specimens. In addition, a thick tributary of the MHV, sometimes arising from S6, did not run along, but penetrated the S5/S6 border plane from the lateral to the medial side. Therefore, the three hepatic veins (RHV, SHV, MHV) often did not aid the identification of the liver segments in the region. Consequently, the less than ideal combinations of irregular configurations of the portal and hepatic venous systems suggest that the right posterior segments cannot be conclusively identified anatomically in 30–40% of cases. Other means of identification, such as the conventional proportional manner (the upper and lower halves of the posterior sector roughly correspond to S6 and S7) may be required. Clin. Anat. 12:229–244, 1999.

Efficacy of SPIO-MR imaging in the diagnosis of liver metastases from colorectal carcinomas.

Aim: To determine whether superparamagnetic iron oxide-enhanced magnetic resonance imaging (SPIO-MRI) could replace intravenous contrast-enhanced spiral CT (iv-CT) and spiral CT during arterial portography (CTAP) combined with spiral CT hepatic angiography (CTHA) in the diagnosis of liver metastases from colorectal carcinomas. Methods: Twenty-six adult patients with liver metastases were studied preoperatively by means of iv-CT, CTAP/CTHA, and SPIO-MRI. Preoperative diagnoses using iv-CT, CTAP/CTHA, and SPIO-MRI were compared with intraoperative and pathological findings in resected specimens. The gold standard for the lesions that were resected was histological examination. Intraoperative findings represented the gold standard for lesions that were not resected. Results: Twenty-six patients were found to have a total number of 43 liver metastases. The sensitivities of iv-CT, CTAP/CTHA, and SPIO-MRI were 74.4, 100, and 90.7%, respectively. SPIO-MRI was significantly superior to iv-CT (p < 0.05). The positive predictive values of iv-CT, CTAP/CTHA, and SPIO-MRI were 97.0, 91.5, and 100%, respectively. CTAP/CTHA yielded four false-positive lesions. In contrast, we detected no false-positive findings using SPIO-MRI. Conclusions: These results suggest that SPIO-MRI might not completely replace CTAP/CTHA, but could replace iv-CT in the diagnosis of liver metastases from colorectal carcinomas. It is thought that SPIO-MRI is a promising imaging modality for diagnosing liver metastases in patients with colorectal carcinoma because of its relatively high sensitivity and extremely high specificity.

Vascular endothelial growth factor C promotes lymph node metastasis in a rectal cancer orthotopic model.

PurposeVascular endothelial growth factor C (VEGF-C), a novel member of the vascular endothelial growth factor family, is a relatively specific lymphangiogenic growth factor. It has been suggested that increased expression of VEGF-C in primary tumors is correlated with lymph node metastasis. We conducted this study to determine whether VEGF-C directly affects lymphangiogenesis and lymph node metastasis in colorectal cancer.MethodsFor an accurate analysis and clear visualization of metastases, the rectal cancer cell line, DLD1, was engineered to stably express green fluorescent protein (GFP) (DLD1/GFP). We implanted DLD1/GFP cells overexpressing VEGF-C orthotopically into the rectal walls of nude mice.ResultsLymph node metastasis was confirmed in all (100%) of the mice bearing DLD1/GFP-VEGF-C tumors, but in only 25% of the mice bearing control tumors. There were more lymph node metastases per mouse in the mice bearing DLD1/GFP-VEGF-C tumors than in the mice bearing control tumors. There were no differences in cell growth and motility in vitro or in the resulting tumor volume from the implanted cells between the two groups. Immunohistochemical staining revealed that VEGF-C induced the growth of lymphatic vessels, which were enlarged in the tumor periphery and contained tumor cell emboli.ConclusionThese results suggest that VEGF-C-induced lymphangiogenesis mediates tumor spread and the formation of lymph node metastasis.

A Novel, Easy, and Safe Technique to Repair a Stoma Prolapse Using a Surgical Stapling Device

A stoma prolapse is one of the late complications and often occurs when the stoma is made in an emergency situation. This complication is not lethal, but causes irritable stoma, skin trouble, and difficulty in stoma care. We herein report the case of a 48-year-old female with an end colostomy that was created as an emergency operation 4 months before. On admission, her colostomy protruded approximately 20 cm from the skin with marked redness, swelling, and erosion; it was impossible to treat manually. We repaired the prolapse successfully in a simple procedure with a Proximate Linear Cutter 100. Briefly, under mild sedation, the instrument was diagonally inserted into the prolapsed stoma and applied twice on both sides. Then, the base of each divided tissue was stapled and cut with the same device. Finally, the prolapse was completely repaired without major bleeding and severe pain. We have applied this novel technique successfully in 5 further cases, and there have been no complications or recurrences. This technique can be performed without spinal or general anesthesia and seems to be a very useful procedure for patients with prolapse of a stoma.

A novel isoform of TUCAN is overexpressed in human cancer tissues and suppresses both caspase-8- and caspase-9-mediated apoptosis.

Caspase-associated recruitment domains (CARD) are protein-protein interaction modules found extensively in proteins that play important roles in apoptosis. One of the CARD-containing proteins, TUCAN (CARD8), was reported previously as an antiapoptotic protein with a molecular weight of 48 kDa, which was up-regulated in colon cancer cells. We identified a novel isoform of TUCAN with a molecular weight of 54 kDa. The new variant of TUCAN, termed TUCAN-54, was expressed in gastric, colon, and breast cancer tissues but was barely detected in normal noncancerous tissues, whereas 48-kDa TUCAN was detected in tumor tissues and noncancerous tissues. To know the function of TUCAN-54 in the apoptosis of cancer cells, TUCAN-54 was overexpressed in tumor cells by gene transfection. Its overexpression inhibited pro-caspase-9 activation, leading to the suppression of the cell death induced by a protein kinase inhibitor, staurosporine, or a chemotherapeutic reagent, etoposide (VP-16). In contrast, specific small interfering RNA-mediated suppression of TUCAN-54 expression in tumor cells increased the VP-16-induced cell death rate, indicating that expression of TUCAN-54 might be associated with chemoresistance of tumor cells. In addition, it inhibited caspase-8 activation as well, thereby suppressing Fas-induced cell death. It was revealed that Fas-associated death domain was physically associated with TUCAN-54 but not with 48-kDa TUCAN. Thus, TUCAN-54 might be a novel tumor-specific antiapoptotic molecule expressed in a variety of human cancer tissues, which might aggravate malignant potential of cancer cells, such as chemoresistance and immunoresistance.

Giant peritoneal loose body in the pelvic cavity: report of a case.

This report describes a giant peritoneal loose body in the pelvic cavity. A 63-year-old man who was asymptomatic underwent a routine medical examination, which revealed a tumor in the pelvic space. Computed tomography and magnetic resonance imaging showed a smooth-surfaced mass with two marked calcifications in the central position. Preoperatively, we suspected a calcified leiomyoma originating from the wall of the sigmoid colon; however, at laparoscopic surgery we extracted a hard, egg-shaped mass 5 cm in diameter, with detached appendices epiploicae. Histological examination revealed that this peritoneal loose body was made up of thick layers of fibrous tissue with a few cellular components, and necrotic fat tissue in the central position. Small peritoneal loose bodies are occasionally found during laparotomy or autopsy, but such a large one is very unusual.