Photodynamic therapy with paclitaxel-encapsulated indocyanine green-modified liposomes for breast cancer.

Background: Photodynamic therapy (PDT) involves the administration of a photosensitizing agent and irradiation of light at an excitation wavelength that damages tumor cells without causing significant damage to normal tissue. We developed indocyanine green (ICG)-modified liposomes in which paclitaxel (PTX) was encapsulated (ICG-Lipo-PTX). ICG-Lipo-PTX accumulates specifically in tumors due to the characteristics of the liposomes. The thermal and photodynamic effects of ICG and the local release of PTX by irradiation are expected to induce not only antitumor effects but also cancer immunity. In this study, we investigated the antitumor effects of ICG-Lipo-PTX in breast cancer. Methods: The antitumor effects of ICG-Lipo-PTX were examined in xenograft model mice subcutaneously implanted with KPL-1 human breast cancer cells. ICG-Lipo-PTX, ICG-Lipo, or saline was administered intraperitoneally, and the fluorescence intensity was measured with a fluorescence imaging system (IVIS). Intratumor temperature, tumor volume, and necrotic area of tumor tissue were also compared. Next, we investigated the induction of cancer immunity in an allogeneic transplantation model in which BALB-MC mouse breast cancer cells were transplanted subcutaneously in the bilateral inguinal region. ICG-Lipo-PTX was administered intraperitoneally, and PDT was performed on only one side. The fluorescence intensity measured by IVIS and the bilateral tumor volumes were compared. Cytokine secretory capacity was also evaluated by ELISPOT assay using splenocytes. Results: In the xenograft model, the fluorescence intensity and temperature during PDT were significantly higher with ICG-Lipo-PTX and ICG-Lipo in tumor areas than in nontumor areas. The fluorescence intensity in the tumor area was reduced to the same level as that in the nonirradiated area after two times of irradiation. Tumor growth was significantly reduced and the percentage of necrotic area in the tumor was higher after PDT in the ICG-Lipo-PTX group than in the other groups. In the allograft model, tumor growth on day 14 in the ICG-Lipo-PTX group was significantly suppressed not only on the PDT side but also on the non-PDT side. In addition, the secretion of interferon-γ and interleukin-2 was enhanced, whereas that of interleukin-10 was suppressed, in the ICG-Lipo-PTX group. Conclusion: The PDT therapy with ICG-Lipo-PTX may be an effective treatment for breast cancer.

Discrepancies Between Morphological and Immunohistochemical Classifications Are Associated With Prognoses and Subtypes of Lung Cancer.

BACKGROUND/AIM: Immunohistochemical (IHC) staining has been routinely used to distinguish adenocarcinoma (ADC) and squamous cell carcinoma (SCC) of the lungs; however, it is challenging to obtain an accurate diagnosis, especially for cases with discrepancies between IHC and hematoxylin and eosin (H&E) staining results. This study aimed to clarify the clinicopathological characteristics of these discrepant cases. PATIENTS AND METHODS: Tissue microarray specimens from 321 patients with ADC and SCC were used for H&E and IHC staining of thyroid transcription factor 1 (TTF-1), Napsin A, cytokeratin 5/6 (CK5/6), p40, and p63. The pathological diagnosis was made based on (1) H&E, (2) IHC, and (3) both H&E and IHC results. Discrepant cases were defined as those with different diagnoses based on the H&E and IHC results. RESULTS: A total of 32 (10%) discrepant cases were identified. ADC (3.9%) showed fewer discrepant cases than SCC (51%). Discrepant cases of ADC had a significantly higher proportion of poorly differentiated tumors and subtypes of solid and invasive mucinous ADC, and they also had shorter overall and disease-free survival than concordant cases. Solid and invasive mucinous ADC cases showed low positivity for TTF-1 (84% and 40%, respectively) and Napsin A (88% and 80%, respectively), and invasive mucinous ADC cases showed high positivity for CK5/6 (80%). The sensitivity and specificity of TTF-1+Napsin A for ADC were 91% and 83%, respectively, whereas those of CK5/6+p40 for SCC cases were 90% and 96%, respectively. CONCLUSION: Discrepant cases of ADC are associated with solid and invasive mucinous subtypes and shorter survival.

Association of longer telomere length in cancer cells and cancer-associated fibroblasts with worse prognosis.

BACKGROUND: Telomere dysfunction has been reported to be directly involved in carcinogenesis owing to chromosomal instability and immortalization; however, the clinicopathological significance of telomeres remains controversial. We have shown that telomere shortening occurs in normal-appearing duct cells at initiation and then continues during the progression of pancreatic cancer. In this study, we determined the clinicopathological and prognostic value of telomere length (TL) in cancer progression. METHODS: TL in both cancer cells and cancer-associated fibroblasts (CAFs) was analyzed by high-throughput quantitative fluorescence in situ hybridization using a previously reported cohort comprising 1434 cases of adenocarcinoma (ADC), squamous cell carcinoma (SCC), adenosquamous carcinoma, hepatocellular carcinoma, and renal cell carcinoma (RCC), which are known cancers with a statistically significantly low incidence of alternative lengthening of telomeres. Cases were divided into 2 groups as follows: longer and shorter telomeres, according to the median TL of cancer cells and CAFs. The statistical significance of TL in cancer cells and CAFs on clinicopathological characteristics and prognosis was analyzed. RESULTS: There was a close association between TL in cancer cells and CAFs. Longer telomeres in cancer cells and CAFs were associated with aggressive features such as advanced stage, high mitosis score and nuclear score, poorly differentiated cancer, and desmoplastic stroma in ADC. Furthermore, a longer TL was an independent prognostic factor for ADC, SCC, and RCC. CONCLUSIONS: Longer telomeres are associated with worse prognosis in ADC, SCC, and RCC. Thus, TL is a novel biomarker for the diagnosis of aggressive cancers with poor prognoses.

Inhibitory effects and gene expression analysis of chemotherapeutic photodynamic therapy by using a liposomally formulated indocyanine green derivative.

BACKGROUND: Photodynamic therapy (PDT) utilizes the enhanced permeability retention effect of photosensitizers and is less invasive and more selective than traditional chemotherapy. We constructed a chemotherapeutic PDT (chemo-PDT) nanoscale drug delivery system using a liposomally formulated indocyanine green derivative (ICG-Lipo) that encapsulated carboplatin and docetaxel (ICG-Lipo-C&D). METHODS: The antitumor effect of chemo-PDT mediated by ICG-Lipo-C&D was evaluated in a murine colon 26 CDF1 mouse model. Gene expression in tumor tissues was analyzed by RNA sequencing. RESULTS: Chemo-PDT using ICG-Lipo-C&D demonstrated an even stronger PDT-enhancing effect than did ICG-Lipo due to the synergistic effect of carboplatin and docetaxel. In addition, gene expression analysis showed that PDT with ICG-Lipo-C&D increased the expression of immune-related genes and decreased the expression of cytoskeleton-related genes. CONCLUSIONS: Chemo-PDT using ICG-Lipo as a photosensitizer as well as a drug delivery system with an enhanced permeability retention effect may be a promising cancer therapy.

Efficacy of Nanofiber Sheets Incorporating Lenvatinib in a Hepatocellular Carcinoma Xenograft Model.

Lenvatinib has a high response rate in unresectable advanced hepatocellular carcinoma (HCC). In this study, we investigated whether lenvatinib-incorporating poly(ε-caprolactone) sheets (lenvatinib sheets) as a drug delivery system (DDS) exerted antitumor effects in a murine HCC model. The lenvatinib sheets were designed for sustained release of approximately 1 mg lenvatinib for 14 days. For 14 days, 1 mg lenvatinib was orally administered to mice. Then, we compared the antitumor effects of lenvatinib sheets with those of oral lenvatinib. The tumor volume, body weight, and serum lenvatinib level were measured for 14 days. A peritoneal dissemination model was established to examine the survival prolongation effect of the lenvatinib sheets. Tumor growth was significantly inhibited in the lenvatinib sheet group compared with that in the no treatment and oral groups. The antitumor effect was significantly higher in the lenvatinib sheet group. Regardless of the insertion site, the serum lenvatinib levels were maintained and showed similar antitumor effects. The mitotic index was significantly inhibited in the lenvatinib sheet group compared with that in the control group. Furthermore, lenvatinib sheets improved the 30-day survival. Lenvatinib sheets showed sufficient antitumor effects and may serve as an effective novel DDS for advanced HCC.

Telomere length was associated with grade and pathological features of meningioma.

Telomeres are tandem repeats of the TTAGGG sequence at chromosomal ends and afford protection against chromosomal instability. To investigate the contribution of telomere dysfunction in meningiomas, here we estimate the associations between telomere length, tumor grade, and proliferation index in a series of 14 archived samples, using quantitative-fluorescence in situ hybridization, Ki67 immunostaining, and pathological analysis. The number of mitoses per 10 high-power fields (HPF) and Ki67 index was higher in grade III cases than in grade I or grade II cases. Telomere length was negatively associated with both the number of mitoses/10HPF and Ki67 index. Meningioma cases with atypical mitosis, a morphological marker of chromosomal instability, exhibited shortened telomeres. Among telomere-shortened meningioma cases, 40% were grade I, 20% were grade II, and 100% were grade III. In grade I or II meningiomas, shortened telomeres lacked high proliferation activity and atypical mitosis. In conclusion, telomere shortening might be pivotal in the development of high-grade meningioma. Analysis of telomere length might be a selective marker for meningiomas with high-grade malignant potential.

A novel nairovirus associated with acute febrile illness in Hokkaido, Japan.

The increasing burden of tick-borne orthonairovirus infections, such as Crimean-Congo hemorrhagic fever, is becoming a global concern for public health. In the present study, we identify a novel orthonairovirus, designated Yezo virus (YEZV), from two patients showing acute febrile illness with thrombocytopenia and leukopenia after tick bite in Hokkaido, Japan, in 2019 and 2020, respectively. YEZV is phylogenetically grouped with Sulina virus detected in Ixodes ricinus ticks in Romania. YEZV infection has been confirmed in seven patients from 2014-2020, four of whom were co-infected with Borrelia spp. Antibodies to YEZV are found in wild deer and raccoons, and YEZV RNAs have been detected in ticks from Hokkaido. In this work, we demonstrate that YEZV is highly likely to be the causative pathogen of febrile illness, representing the first report of an endemic infection associated with an orthonairovirus potentially transmitted by ticks in Japan.

[Radiation-Induced Breast Angiosarcoma after Conservative Surgery for Breast Cancer].

We present the case of an 88-year-old woman who had undergone breast conserving surgery for left breast cancer 8 years ago.She received postoperative radiotherapy(total dose of 60 G/30 Fr)to the residual breast together with endocrine therapy.She underwent skin biopsy after having had a red skin tumor in the left breast.Angiosarcoma was diagnosed and chemotherapy and radiotherapy were initiated.The patient is alive without recurrence 8 months after chemotherapy.

Characterization of tick-borne encephalitis virus isolated from a tick in central Hokkaido in 2017.

Tick-borne encephalitis virus (TBEV) is a zoonotic virus in the genus Flavivirus, family Flaviviridae. TBEV is widely distributed in northern regions of the Eurasian continent, including Japan, and causes severe encephalitis in humans. Tick-borne encephalitis (TBE) was recently reported in central Hokkaido, and wild animals with anti-TBEV antibodies were detected over a wide area of Hokkaido, although TBEV was only isolated in southern Hokkaido. In this study, we conducted a survey of ticks to isolate TBEV in central Hokkaido. One strain, designated Sapporo-17-Io1, was isolated from ticks (Ixodes ovatus) collected in Sapporo city. Sequence analysis revealed that the isolated strain belonged to the Far Eastern subtype of TBEV and was classified in a different subcluster from Oshima 5-10, which had previously been isolated in southern Hokkaido. Sapporo-17-Io1 showed similar growth properties to those of Oshima 5-10 in cultured cells and mouse brains. The mortality rate of mice infected intracerebrally with each virus was similar, but the survival time of mice inoculated with Sapporo-17-Io1 was significantly longer than that of mice inoculated with Oshima 5-10. These results indicate that the neurovirulence of Sapporo-17-Io1 was lower than that of Oshima 5-10. Using an infectious cDNA clone, the replacement of genes encoding non-structural genes from Oshima 5-10 with those from Sapporo-17-Io1 attenuated the neuropathogenicity of the cloned viruses. This result indicated that the non-structural proteins determine the neurovirulence of these two strains. Our results provide important insights for evaluating epidemiological risk in TBE-endemic areas of Hokkaido.

[A Case of Pulmonary Tumor Thrombotic Microangiopathy(PTTM)Diagnosed During Chemotherapy for Metastatic Breast Cancer].

A 46-year-old woman with metastatic breast cancer developed dyspnea that progressed relatively rapidly during chemotherapy. Chest-abdominal CT revealed wedge-shaped infiltration shadow, and cardiac catheterization revealed elevated pulmonary artery pressure. Aspiration cytology of pulmonary arterial blood was performed and malignant cells were confirmed. Chemotherapy was difficult to continue because of deterioration in general condition, and she died 7 days after diagnosis. This time, we report a case of PTTM for which pulmonary arterial blood cytology was useful for diagnosis.

[Temporary Home Management of a Refractory Pleural Effusion with an Indwelling Catheter].

BACKGROUND: Pleurocentesis and pleurodesis are the common treatments for pleural effusion. However, most home-visit physicians usually hesitate to perform these treatments for patients confined at home. CASE: A 51-year-old woman developed breast cancer(ER+, HER2-)at the age of 39 years. She underwent a partial mastectomy of the right breast. Nine years later, metastatictumors in the lungs, and hilar and mediastinal lymph nodes were found. The patient was admitted to our hospital because of the progression of pleural effusion and dyspnea. On the day of admission, the aspiration catheter was placed in the left lung with continuous suction, but pleurodesis could not be performed as the left lung did not re-expand enough. As the patient requested to go home as soon as possible, she was discharged with the catheter in place. Three days after the discharge, the home-visit physician drained 340 mL of fluid through the catheter. Six days after the discharge, the patient was readmitted to the hospital with malaise and dyspnea, but no signs of complications associated with the indwelling catheter use were observed. The patient died 4 days after the readmission. CONCLUSION: This case suggests that draining fluid using an indwelling pleural catheter as a home-based healthcare measure is one of the simplest and safest options.

[Abnormal Nipple Discharge].

BACKGROUND: Breast carcinoma and precancer are thought to start in the lining of the milk duct or lobule. Ductography and fiberoptic ductoscopy have beenadvocated as the mainprocedures inpatien ts with nipple discharge. METHODS: We investigated the usefulness of microdochectomy(MD)by using indocyanine green(ICG)fluorescence imaging. ICG and indigo carmine were injected into the mammary duct. A periareolar incision was made, and a fluorescence image of the demarcated mammary duct segment was obtained. CONCLUSION: MD using indocyanine green fluorescence imaging is a useful procedure in guiding subsequent breast surgery in the treatment of nipple discharge.

[A Case of Ductal Carcinoma In Situ(DCIS)Diagnosed by Excisional Biopsy with Benign Cytology Findings of Fine-Needle Aspiration].

Nipple discharge is a common symptom and frequently results from benign tumors. However, there is a 5-30% risk of malignancy. A 65-year-old woman presented at the hospital because of bloody nipple discharge in her right breast. She had noticed an abnormal nipple discharge for several months. Mammography showed focal asymmetric densities without calcification in the middle outer quadrant of her right breast. Ultrasonography indicated a 1.5×1.1 cm sized cyst with fluid-fluid level. Breast MRI showed a simple cyst with a benign contrast enhancement pattern. No malignant cells were observed by fine-needle aspiration. Considering the low sensitivity of mammography and breast MRI to DCIS, we performed an excisional biopsy. Histological examination revealed that the lesion was DCIS. The patient underwent right total mastectomy and was diagnosed with low grade DCIS(ER-positive, PgR-positive, HER2-negative). She continues endocrine therapy with an aromatase inhibitor.

Serologic Evidence of Tick-Borne Encephalitis Virus Infection in a Patient with Suspected Lyme Disease in Japan.

Tick-borne encephalitis (TBE) is widely prevalent on the Eurasian continent, including Japan, but four cases of TBE have been reported in Japan. To inspect unconfirmed TBE cases in Japan, we conducted a retrospective seroepidemiological study of a total of 158 samples from 81 meningoencephalitis patients suspected as Lyme disease. Two serum samples from one patient showed neutralizing antibodies against TBE virus. The patient with severe and progressive encephalitis had a history of tick bite in Hokkaido in 2012. These results demonstrated that tick-borne encephalitis virus (TBEV) case was actually unconfirmed in Japan. Further seroepidemiological surveys are required to identify unconfirmed TBEV infections to consider the pros and cons of introducing specific countermeasures including vaccination in Japan.

[A Successful Treatment of Locally Advanced Breast Cancer with Using Mohs' Paste and Chemotherapy - A Case Report].

Foul smell and large amounts ofexudate, bleeding are the most common and serious symptoms with locally advanced breast cancer(LABC). Mohs' paste is made ofa mixture ofzinc chloride and used for treatment ofmalignant skin tumors. Recently some reports show that Mohs' paste is useful for treatment of malignant tumor including unresectable breast cancer and skin metastasis ofcancer. Mohs' paste is useful for reducing symptoms such as foul smell and exudate, Bleeding. We report a successful case of treatment for LABC with using Mohs' paste and chemotherapy and surgery.

[Local Control by Mastectomy in Advanced Breast Cancer with Liver Metastasis after Chemotherapy, Radiotherapy, and Hyperthermia - A Case Report].

Advanced breast cancer has a poor prognosis compared to early breast cancer; however, quality of life and radical operation can be improved in some case by using multidisciplinary treatment. A 54-year-old woman was examined at the hospital because of an enlarging tumor in the left breast. She was aware of a lump for 3 years. Results of the initial examination indicated invasive ductal carcinoma with liver metastasis. She first received chemotherapy(AC followed by weekly paclitaxel). After 4 courses of weekly paclitaxel, computed tomography revealed axillary lymph nodes involved in the axillary vein. Operation was difficult and conversion therapy was administered. The patient underwent radiotherapy, hyperthermia, and hormone therapy. After 1 year from the start of hormone therapy, the metastasis had disappeared and the patient underwent operation in our unit. Eight months after operation, no recurrence was observed.

[Evaluation of the Small Excision Using a Sonographically Visible Breast Biopsy Marker(HydroMARK®)].

In case an operation is necessary after an ST-VAB for microcalcification, a wide excision needs to be performed because of the loss of marking. The HydroMARK®breast biopsy marker can be visualized using an ultrasound or mammography and facilitates a small excision. Six months after the deployment, all markers were visualized using an ultrasound. Thus, Hydro MARK®can decrease the sample size(11 g)in open biopsies and correctly diagnose ADH. However, no HydroMARK®cases were diagnosed with ADH in large samples(44 g, 32 g). Five malignant histology cases underwent mastectomy, and the distance between the HydroMARK®and tumor was 300 mm. HydroMARK®appears to be a safe and effective marker after a stereotactic biopsy for calcification, which facilitates an exact small excision of lesion surgically.

[Radiotherapy and Eribulin Mesylate for Mediastinal Node Metastasis from Breast Cancer-A Case Report].

A69 -year-old woman was diagnosed as having local advanced breast cancer. She was treated with neoadjuvant chemotherapy( AC followed by paclitaxel)and followed up with left total mastectomy with axillary lymphadenectomy for breast cancer(pT3N1aM0, Stage ⅢA, ER positive, PgR positive, and HER2 negative). She received adjuvant therapy with chest wall irradiation and an aromatase inhibitor. Two years after the mastectomy, mediastinal lymph node and rib metastases and dissemination appeared. We changed the regimen to capecitabine. She continued the capecitabine therapy for 7 years and was found to have multiple lung metastasis. Therefore, we chose eribulin mesylate therapy. Ten days after eribulin mesylate (1.0mg/body)was first administered, she suddenly had difficulty breathing, and chest CT revealed left low lobe atelectasis. The superior mediastinal lymph node had grown rapidly and compressed the left main bronchus and superior vena cava. To reduce the lymph node size, we started radiotherapy(50 Gy/25 Fr)for the superior mediastinal area in addition to the eribulin mesylate therapy. After the radiotherapy, chest CT revealed a remarkable reduction of lymph node size and full pulmonary reexpansion. One year after the radiotherapy, she is continuing treatment without systemic progression.

[Examination of Stereotactic Mammotome Biopsy for Microcalcification in Our Hospital].

We introduced stereotactic mammotome biopsy(ST-MMT)for the purpose of screening and other institutions. There are many benign cases to be diagnosed by pathological findings, so it is thought to be necessary to examine the adaptation of STMMT again. We examined the performance of ST-MMT in a case of a non-palpating calcification lesion. Between August 2013 and December 2016, ST-MMT biopsies were performed for 247 microcalcified lesions revealed by mammography(in both breasts in 9 patients; twice in the ipsilateral breast in 2 patients). The mean age of all patients was 46 years(range, 24- 89 years). We found 39 cases(15.8%)of breast cancer. A final diagnosis of breast cancer was made in 39 patients, who comprised 0% of those with Category 2, 53.8% of those with Category 3, 35.9% of those with Category 4, and 10.3% of those with Category 5. Regarding the morphology and distribution of microcalcifications, breast cancer accounted for 46.2%, 5.1%, 2.6%, 35.9%, 7.7%, and 2.6% of the cases with small round/clustered, amorphous/clustered, pleomorphic/clus- tered, pleomorphic/linear segmental, and fine linear/clustered patterns, respectively. Also, we examined each of the patients, (1) who underwent mammography for medical examinations, (2) who underwent mammography performed at other institutions, (3) who underwent follow-up for microcalcifications and postoperative follow-up mammography. The proportions of breast cancer diagnoses were (1) 11.4%, (2) 20.6%, and (3) 7.1%. Proportions of Category 3 breast cancer were (1) 10.3%, (2) 38.5%, and (3) 5.1%. Among the cases in which ST-MMT was performed in this study, Category 3 accounted for more than half. However, 10.9%(21/192 lesions)were diagnosed as malignant in Category 3. The diagnosis of breast cancer in pa- tients who underwent mammography performed at other institutions was not observed in 79.4%(104/131 lesions), and among the 104 lesions, as a result of reassessment of calcification in our hospital, Category 2 was also included. Calcification in Category 2 lesions was benign in all cases. It was suggested that the indication for ST-MMT biopsy could be further narrowed down by being careful not to over-diagnose.