Survival analysis of multiple peptide vaccination for the selection of correlated peptides in urological cancers.

Peptide-based cancer vaccines are able to induce strong immune responses, but their clinical results are unsatisfactory. To determine clinically correlated peptides, we analyzed survival data from urological cancer patients treated by personalized peptide vaccination (PPV), in which different multiple peptides were used for individual patients based on human leukocyte antigen (HLA) type and pre-existing immunity. Survival data were obtained from a database of 265 urological cancer patients treated in 5 clinical PPV trials comprising 154 patients with castration-resistant prostate cancer (CRPC) and 111 patients with advanced urothelial cancer (UC). Expression of tumor-associated antigens (TAA) was evaluated in 10 prostate cancer tissues, 4 metastatic lymph nodes from prostate cancer, and 10 UC tissues using immunohistochemical staining. Clinical efficacy of individual peptides for overall survival was evaluated by the Cox proportional hazards regression model. All TAA coding candidate peptides used in PPV treatment were expressed in tumor cells from prostate cancer and UC samples except for p56Lck in both, and prostate-specific antigen (PSA), prostatic acid phosphatase (PAP) and prostate-specific membrane antigen (PSMA) in the UC samples. Patients with the following peptides had a significantly longer survival than patients without the peptides (hazard ratio <1.0, 95% confidence intervals <1.0 and P < .05): SART3-109, PTHrP-102, HNPRL-140, SART3-302 and Lck-90 in CRPC patients, and EGF-R-800, Lck-486, PSMA-624, CypB-129 and SART3-734 in advanced UC patients, respectively. Correlated peptides selected using both survival data and pre-existing immunity for PPV treatment may enhance the clinical benefits for urological cancer patients.

TRPA1-dependent reversible opening of tight junction by natural compounds with an α,ß-unsaturated moiety and capsaicin.

The delivery of hydrophilic macromolecules runs into difficulties such as penetration of the cell membrane lipid bilayer. Our prior experiment demonstrated that capsaicin induces the reversible opening of tight junctions (TJs) and enhances the delivery of hydrophilic macromolecules through a paracellular route. Herein, we screened paracellular permeability enhancers other than capsaicin. As TJ opening by capsaicin is associated with Ca2+ influx, we first screened the compounds that induce Ca2+ influx in layered MDCK II cells, and then we determined the compounds' abilities to open TJs. Our results identified several natural compounds with α,ß-unsaturated moiety. A structure-activity relationship (SAR) analysis and the results of pretreatment with reducing reagent DTT suggested the importance of α,ß-unsaturated moiety. We also examined the underlying mechanisms, and our findings suggest that the actin reorganization seen in capsaicin treatment is important for the reversibility of TJ opening. Furthermore, our analyses revealed that TRPA1 is involved in the Ca2+ influx and TJ permeability increase not only by an α,ß-unsaturated compound but also by capsaicin. Our results indicate that the α,ß-unsaturated moiety can be a potent pharmacophore for TJ opening.

Immunological efficacy of herbal medicines in prostate cancer patients treated by personalized peptide vaccine.

This randomized phase II study investigated the immunological efficacy of herbal medicines (HM) using Hochu-ekki-to and Keishi-bukuryo-gan in combination with personalized peptide vaccination (PPV) for castration-resistant prostate cancer (CRPC). Seventy patients with CRPC were assigned to two arms; PPV plus HM or PPV alone. Two to four peptides were chosen from 31 peptides derived from cancer antigens for a s.c. injection of PPV given eight times according to the patient's human leukocyte antigen type and levels of antigen-specific IgG titer before PPV treatment. Peptide-specific CTL, IgG, regulatory T cells (Treg), monocytic myeloid-derived suppressor cells (Mo-MDSC), and interleukin-6 (IL-6) responses were measured before and at the eighth vaccination. Clinical outcomes were also analyzed. Combination therapy of PPV with HM was well tolerated without severe adverse events. There was no significant change in antigen-specific IgG, CTL, Treg or clinical outcomes. Combination therapy of PPV with HM stabilized the frequency of Mo-MDSC (1.91%-1.92%, P = 0.96) and serum levels of IL-6 (19.2 pg/mL to 16.1 pg/mL, P = 0.63) during the treatment. In contrast, the frequency of Mo-MDSC and levels of IL-6 in the PPV-alone group were significantly increased (0.91%-1.49% for Mo-MDSC and 9.2 pg/mL to 19.4 pg/mL for IL-6, respectively). These results suggest that the combined use of HM has the potential to prevent the immunosuppression induced by Mo-MDSC or IL-6 during immunotherapy. More research is needed to validate the findings of the present study.

Personalized peptide vaccination as second-line treatment for metastatic upper tract urothelial carcinoma.

This study investigated the applicability of personalized peptide vaccination (PPV) for patients with metastatic upper tract urothelial cancer (mUTUC) after failure of platinum-based chemotherapy. In this single arm, open-label, phase II clinical trial, patients with mUTUC received PPV at a single institution. Personalized peptide vaccination treatment used a maximum of four peptides chosen from 27 candidate peptides according to human leukocyte antigen types and peptide-reactive IgG titers, for six s.c. injections weekly as one cycle. The safety of PPV, as well as its influence on host immunity and effect on overall survival were assessed. Forty-eight patients were enrolled in this study. Personalized peptide vaccinations were well tolerated without severe adverse events. Median survival time was 7.3 months (95% confidence interval [CI], 5.3-13.1) with 13.0 months for patients receiving combined salvage chemotherapy (95% CI, 5.7-17.5) and 4.5 months for patients receiving PPV alone (95% CI, 1.7-10.1) (P = 0.080). Patients with positive CTL responses showed a significantly longer survival than patients with negative CTL responses (hazard ratio, 0.37; 95% CI, 0.16-0.85; P = 0.019). Multivariate Cox regression analysis showed that lower numbers of Bellmunt risk factors and lower levels of B-cell activating factor were significantly associated with favorable overall survival for patients under PPV treatment. This study indicated that PPV for patients with mUTUC after failure of platinum-based chemotherapy induced substantial peptide-specific CTL responses without severe adverse events and has the potential to prolong survival when combined with salvage chemotherapy. UMIN Clinical Trials Registry ID: 000001854.

Clinical development of immunotherapy for prostate cancer.

Prostate cancer is the most common cancer in men, and the second leading cause of cancer-related death in Western countries. Prostate cancer-related death occurs in patients with metastatic castration-resistant prostate cancer. Although several new drugs for castration-resistant prostate cancer have been approved, each of these has prolonged survival by just a few months. Consequently, new therapies are sorely needed. Recently, it has been recognized that immunotherapy is an effective treatment for prostate cancer patients. Several strategies, such as cancer vaccines and immune checkpoint inhibitors, have been investigated in clinical studies for prostate cancer patients. In the present review, the results of the most recent clinical studies investigating immunotherapy in prostate cancer patients are reported, and the future clinical development of immunotherapy for prostate cancer is discussed.

A randomized phase II clinical trial of personalized peptide vaccination with metronomic low-dose cyclophosphamide in patients with metastatic castration-resistant prostate cancer.

This study investigated the effect of metronomic cyclophosphamide (CPA) in combination with personalized peptide vaccination (PPV) on regulatory T cells (Treg) and myeloid-derived suppressor cells (MDSC), and whether it could improve the antitumor effect of PPV. Seventy patients with metastatic castration-resistant prostate cancer were randomly assigned (1:1) to receive PPV plus oral low-dose CPA (50 mg/day), or PPV alone. PPV treatment used a maximum of four peptides chosen from 31 pooled peptides according to human leukocyte antigen types and antigen-specific humoral immune responses before PPV, for 8 subcutaneous weekly injections. Peptide-specific cytotoxic T lymphocyte (CTL) and immunoglobulin G responses were measured before and after PPV. The incidence of grade 3 or 4 hematologic adverse events was higher in the PPV plus CPA arm than in the PPV alone arm. Decrease in Treg and increase in MDSC were more pronounced in PPV plus CPA treatment than in PPV alone (p = 0.036 and p = 0.048, respectively). There was no correlation between the changes in Treg or MDSC and CTL response. There was no difference in positive immune responses between the two arms, although overall survival in patients with positive immune responses was longer than in those with negative immune responses (p = 0.001). Significant differences in neither progression-free survival nor overall survival were observed between the two arms. Low-dose CPA showed no change in the antitumor effect of PPV, possibly due to the simultaneous decrease in Treg and increase in MDSC, in patients under PPV.

Fabrication of interconnected porous calcite by bridging calcite granules with dicalcium phosphate dihydrate and their histological evaluation.

Interconnected porous calcite has attracted attention as an artificial bone replacement material and as a precursor for the fabrication of carbonate apatite, which is also an artificial bone replacement material. In this study, calcite granules were exposed to acidic calcium phosphate solution, and the feasibility of fabricating interconnected porous calcite using this process was evaluated. No setting reaction was observed under the nonloading condition. In contrast, under loading conditions, calcite granules were bridged with dicalcium phosphate dihydrate crystals, and the calcite granules set into interconnected porous calcite foam. When applied 0.4 MPa of loading pressure during sample preparation, compressive strength of the obtained interconnected porous calcite was ∼1.5 MPa. The exposure of the calcite granules to acidic calcium phosphate solution under loading conditions was the key for the setting reaction to occur. This is because calcite granules cannot contact one another under the nonloading condition because of bubble formation on the surfaces of the calcite granules. The interconnected porous calcite revealed excellent tissue response, and new bone was able to penetrate into the porous calcite 2 weeks after implantation. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A 104A: 652-658, 2016.

Immunotherapy in prostate cancer: challenges and opportunities.

Although treatment options for castration-resistant prostate cancer (CRPC) have increased over the last decade, there remains a need for strategies that can provide durable disease control and long-term benefit. Recently, immunotherapy has emerged as a viable and attractive strategy for the treatment of CRPC. To date, there are multiple strategies to target the immune system, and several approaches including therapeutic cancer vaccines and immune checkpoint inhibitors have been most successful in clinical trials. With regard to this, we report the results of the most recent clinical trials investigating immunotherapy in CRPC and discuss the future development of immunotherapy for CRPC, as well as the potential importance of biomarkers in the future progress of this field.

[Peptide vaccination for castration-resistant prostate cancer].

Since both tumor cells and host immune cell repertoires are diverse and heterogeneous, immune responses against tumor associated antigens shall be substantially different among individual patients with prostate cancer. Subsequently, selection of suitable peptide vaccines for individual patients based on the pre-existing host immunity before vaccination could induce potent anti-tumor responses capable of providing clinical benefit for prostate cancer patients. We have developed a novel immunotherapeutic approach of personalized peptide vaccination (PPV) in which a maximum of four human leukocyte antigen (HLA)-class IA-matched peptides were selected for vaccination among pooled peptides based on both HLA-class IA type and the pre-existing host immunity before vaccination. We discuss our recent results of clinical studies of peptide vaccination for castration-resistant prostate cancer and the future direction of therapeutic cancer vaccines.

A case of primary mucosa-associated lymphoid tissue lymphoma of the prostate.

We report a case of primary mucosa-associated lymphoid tissue (MALT) lymphoma of the prostate. A 67-year-old man presented with urinary obstruction and an elevated prostate-specific antigen (PSA) level. A physical examination revealed mild prostate enlargement and no lymphadenopathy. A needle biopsy and immunohistochemical studies of the prostate were performed, which revealed marginal zone B-cell MALT-type lymphoma. A bone marrow aspiration and biopsy did not show involvement by lymphoma. Magnetic resonance imaging (MRI) of the abdomen and the pelvis revealed no lymphadenopathy or ascites. There was no involvement of other sites by lymphoma. The patient was diagnosed and staged as extranodal marginal zone B-cell MALT-type lymphoma of the prostate, low grade and stage I. The patient received external beam radiation therapy to the prostate with a total dose of 3600cGy in 22 fractions, and became free of disease within the following 15 months.

[Fisher syndrome with optic neuropathy].

UNLABELLED: Fisher syndrome with optic neuropathy has been rarely reported. We report a 78-year-old Frenchman with Fisher syndrome. The patient complained of dizziness and bilateral blurred vision. His corrected visual acuity was 0.03 in the right eye, and 0.02 in the left eye. Deep tendon reflexes were absent. A few days later, bilateral complete external ophthalmoplegia appeared. Both pupils were dilated and pupillary reflexes were absent. Serum anti-GQlb antibodies and anti-GT1a antibodies were detected. After intravenous immunoglobulin treatments, all neurological symptoms including optic neuropathy and external ophthalmoplegia disappered except for pupillary dilatation associated with light-near dissociation. Tonic pupil indicated disorder of the peripheral nervous system. CONCLUSION: Fisher syndrome may complicate optic neuropathy.

Anti-stress, anti-HIV and vitamin C-synergized radical scavenging activity of mulberry juice fractions.

Anti-stress and anti-HIV activity of mulberry juice were separated by centrifugation. The anti-stress activity was enriched in the supernatant fraction whereas the anti-HIV activity in the precipitate fraction. Oral administration of the supernatant fraction significantly reduced the elevated plasma level of lipid peroxide in mice loaded with water immersion restraint stress. The kinetic study revealed that the anti-stress activity was maintained for 4 hours after cessation of the administration of mulberry juice. The lignin fraction in the precipitate fraction scavenged superoxide and hydroxyl radicals more efficiently than other fractions, in a synergistic fashion with sodium ascorbate. Anti-HIV activity of mulberry juice was concentrated in the lignin fraction, whereas blueberry juice, which has no precipitating fibrous materials, did not show anti-HIV activity. The present study suggests the functionality of mulberry juice as an alternative medicine.

Abducens nerve palsy and postganglionic Horner syndrome with or without severe headache.

OBJECTIVE: To report the clinical features of 9 patients with both abducens nerve palsy and postganglionic Horner syndrome. PATIENTS AND METHODS: Nine patients with this symptom combination were examined by our Neuro-ophthalmology Clinic in Nihon University Itabashi Hospital between 1978 and 2004. Patient ages ranged from 28 to 63 years (average 47.2+/-8.7 years). Six patients were males and 3 were females. Primary diseases, accompanying symptoms and prognoses were surveyed. RESULTS: In primary diseases, neoplasm in the cavernous sinus was observed in 2 patients, sphenoidal sinus cyst in 2, intra-cavernous carotid aneurysm in 2, epipharynx carcinoma in 1, chordoma in the base of the skull in 1, and meningioma in the middle cranial fossa in 1. Five patients with extra-cavernous sinus lesions; sphenoidal sinus cyst, epipharynx carcinoma, chordoma and meningioma, complained of severe headache. However, in patients without severe headache, intra-cavernous sinus lesions such as carotid aneurysm and metastatic carcinoma were detected. After therapy, abducens nerve palsy improved in 5 patients, however, Horner syndrome persisted in all patients. CONCLUSION: We emphasize that this symptom combination is an important sign of lesions in the posterior portion of the cavernous sinus or in its vicinity. Moreover, the presence or absence of severe headache depends on whether the lesion is in the intra-cavernous or extra-cavernous sinus.

[Clinical evaluation of primary position downbeat nystagmus].

PURPOSE: Primary position downbeat nystagmus is a vertical nystagmus with a rapid phase downwards presenting in any gaze. We evaluated causative lesions, etiology, and incidence by age groups. MATERIALS AND METHODS: Twenty-five patients with primary position downbeat nystagmus were enrolled in this study. Their causative lesions, etiology, and frequency by age group were investigated. We divided the patients into 3 groups depending on age; under 30, from 30 to 60, and over 60 years. RESULT: Cerebrovascular disorder was diagnosed in 5 patients. Four patients had cerebellar degeneration. Three patients had multiple sclerosis or Arnold-Chiari malformation. Two patients had cerebellar tumor. In the young group under 30, etiology was mainly cerebellar tumor or multiple sclerosis. Especially cerebellar tumor was limited to this group. In the middle age group, there was no distinctive tendency. In the older group over 60 years, cerebrovascular disorder was predominant. In 10 patients with intracranial lesions, the foci were located in the cerebellum, pons, or medulla oblongata. CONCLUSION: The causes of primary position down beat nystagmus tended to be age-dependent. The locations of causative lesions were the cerebellum and the inferior brain stem.

[Clinical evaluation of primary position upbeat nystagmus].

BACKGROUND: Primary position upbeat nystagmus is a vertical nystagmus presenting in the primary position. However, cases of primary position upbeat nystagmus have rarely been reported. SUBJECTS: We evaluated the locations of lesions and pathogenesis in 11 patients with primary position upbeat nystagmus. The lesions were located mostly in the median portion of the cerebellum and medulla oblongata. In 7 out of 11 patients, intracranial lesions were identified by computed tomography and magnetic resonance imaging. They were located in the median portion of the cerebellum and medulla oblongata except for one adult patient with astrocytoma. The etiologies were 4 tumors, 3 intracranial vascular disorders, 2 Wernicke encephalopathies, 1 congenital anomaly, and 1 inflammation. CONCLUSION: The lesions were located mostly in the median portion of the cerebellum and medulla oblongata, and especially in the cerebellum in children. The main etiologies were tumors in children and vascular disorders and Wernicke encephalopathy in adults.