Anticancer effect and immune induction by hyperthermia of malignant melanoma using magnetite cationic liposomes

The hyperthermic effect of magnetic particles was examined in an in vivo study of mouse B16 melanoma. Magnetite cationic liposomes (MCLs) have a positive surface charge and generate heat under an alternating magnetic field (AMF) by hysteresis loss. MCLs were injected into melanoma nodules, which were then subjected to an AMF. The mice were divided into four groups: group I (control), group II (hyperthermia at 43 degrees C for 30 min, once), group III (hyperthermia at 46 degrees C for 30 min, once) and group IV (hyperthermia at 46 degrees C for 30 min, twice). Complete tumour regression was observed in 90% of the mice in group IV, while no mice in groups I and II and only 40% in group III showed regression. To examine whether hyperthermia caused immune induction in B16 melanoma, in vitro cytotoxicity assays and rechallenge experiments were performed. Cytotoxic activity was observed in the spleen cells of the cured mice in group IV. In the rechallenge experiment, 66% of the cured mice rejected melanoma cells. These results suggest that hyperthermia using MCLs is an effective therapy for melanoma, since this treatment can kill the tumour cells not only by heat but also by inducing an immune response.     

Treatment of disseminated malignant melanoma with cisplatin in combination with whole-body hyperthermia and doxorubicin

Twenty-three patients with disseminated malignant melanoma were entered in a pilot study conducted from January 1983 to January 1988. There were 14 men and nine women. Fifteen patients were available for final evaluation. The criterion for evaluation was the completion of the two thermochemotherapy cycles. Patients received multiple-drug chemotherapy consisting of cisplatin (80 mg/m2, i.v., day 1) and doxorubicin (50 mg/m2, i.v., day 2); applications were repeated on days 22-29. Cisplatin was administered concurrently with whole-body hyperthermia (WBH) on day 1 (body core temperature 41 +/- 0.5 degrees C/h). The following response rates were achieved: complete remission (CR) 0/15; partial remission (PR) 3/15; minor response (MR) 4/15, no change (NC) 3/15; progressive disease (PD) 5/15. These findings are within the range known from the literature of the response to multiple-drug chemotherapy without hyperthermia. Toxicity, in particular renal toxicity, was not enhanced by the combined application of cisplatin and hyperthermia, with the exception of the bone marrow toxicity, which seemed to be slightly more pronounced than that known from normothermic application. Concurrent administration of thermochemotherapy with cisplatin and WBH (41 +/- 0.5 degrees C/h) is no more effective than the multiple-drug chemotherapy without hyperthermia as reported in the literature.     

Treatment of disseminated malignant melanoma with cisplatin in combination with whole-body hyperthermia and doxorubicin

Twenty-three patients with disseminated malignant melanoma were entered in a pilot study conducted from January 1983 to January 1988. There were 14 men and nine women. Fifteen patients were available for final evaluation. The criterion for evaluation was the completion of the two thermochemotherapy cycles. Patients received multipledrug chemotherapy consisting of cisplatin (80 mg/m², i.v., day 1) and doxorubicin (50 mg/m², i.v., day 2); applications were repeated on days 22–29. Cisplatin was administered concurrently with whole-body hyperthermia (WBH) on day 1 (body core temperature 41±0·5°C/h). The following response rates were achieved: complete remission (CR) 0/15; partial remission (PR) 3/15; minor response (MR) 4/15, no change (NC) 3/15; progressive disease (PD) 5/15. These findings are within the range known from the literature of the response to multiple-drug chemotherapy without hyperthermia. Toxicity, in particular renal toxicity, was not enhanced by the combined application of cisplatin and hyperthermia, with the exception of the bone marrow toxicity, which seemed to be slightly more pronounced than that known from normothermic application. Concurrent administration of thermochemotherapy with cisplatin and WBH (41±0·5°C/h) is no more effective than the multiple-drug chemotherapy without hyperthermia as reported in the literature.     

A combination chemotherapy of ACNU and DTIC for advanced malignant melanoma

Fourteen patients with advanced malignant melanoma were treated with a combination chemotherapy consisting of ACNU 100 mg/m2 i.v. on Day 1 in 6 week intervals and DTIC 200 mg/m2 i.v. on Days 1 to 5 at 3 week intervals. Four patients had prior chemotherapy and 2 had prior immunotherapy. Excluding 4 patients received the regimen for adjuvant chemotherapy, 10 of 14 patients were evaluable for response. There were 3 patients of partial responses, 3 minor responses, 1 no change, and 3 progressive diseases. The durations of partial responses were 1, 1, and 8 months, respectively, while the survival times in these patients were 5, 21, and 10 months, respectively. Leukopenia less than 4,000/cmm occurred in 10 of 14 patients (71%) and thrombocytopenia less than 100 X 10(3)/cmm in 9 of 14 patients (64%), moreover, these hematologic toxicities were cumulative. Serum GOT and GPT elevated to 3,460 mu/ml and 1,365 mu/ml, respectively in one patient, but this returned to a normal level one month later. Nausea and vomiting were mild to severe in 12 of 14 patients, being most marked on Day 1 and decreasing intensity during the next several days. Other non-hematologic toxicities including skin rash, fever, and phlebitis were noted in each one patient, respectively. Hematologic toxicity of this regimen was a dose limiting toxicity; therefore, intensive supportive therapy to prevent infection and hemorrhage is essential for the management of the patients during this chemotherapy.     

Prognostic factors in metastatic malignant melanoma treated with combined radiation therapy and hyperthermia

From May 1981 to September 1991, 38 patients with metastatic malignant melanoma were treated with combined radiation therapy and hyperthermia to a total of 97 hyperthermia treatment fields. Prior treatments to these sites included surgery (31 patients, 76 fields), chemotherapy (18 patients, 54 fields), immunotherapy (14 patients, 42 fields) and radiation therapy (7 patients, 13 hyperthermia fields). Hyperthermia was given to fields located in the head and neck region, trunk and extremities in 30, 45 and 22 cases, respectively. Nodular-diffuse tumours were present in 86 fields while 11 fields were treated for microscopic residual tumour deposits. Concurrent radiation therapy was given in 180–400 cGy per fraction, 2–5 times per week for a mean total dose of 4098 cGy per field. Hyperthermia treatments were delivered using either microwave or ultrasound devices (286 and 48 treatments, respectively) with a mean (range) of 3. 4 (1–14) hyperthermia treatments per field for a mean (range) of 43 (10–70) min per field. Patients (n = 34; 84 fields) were available for follow-up for a mean (range) of 14–6 (0–4-82.5) months. At 3 weeks post-treatment, 34 fields had complete, ongoing, or partial responses; 39 fields had no response; and there were no recurrences in the 11 fields treated for microscopic residual disease. Local control was maintained in 31% (26/84) fields with a mean follow-up of 14.6 months. At 36 months, five patients remained alive with complete control of their treated local disease. Statistical analyses revealed that patients with soft tissue metastases only, who were older at the time of hyperthermia, had a longer time between initial diagnosis and hyperthermia treatment, received a higher dose of radiation, had no previous chemotherapy, and had small tumour volumes, had a higher initial response. Multivariate analyses revealed that the three-covariate model including time interval between initial diagnosis and hyperthermia treatment, previous chemotherapy, and metastases to soft tissue only, best predicted response. The results of the investigation support the continued study of combined radiation therapy and hyperthermia treatments for selected patients with metastatic melanoma, and indicate that long-term survival can occasionally be obtained with this approach.     

Growth inhibition of human malignant melanoma transfected with the human interferon-beta gene by means of cationic liposomes

Among the various types of human interferons, human interferon-beta (HuIFNbeta) has the strongest anti-proliferative activity against human melanoma cell lines. Therefore, we investigated the growth inhibitory effect of a cationic liposome containing the HuIFNbeta gene on human melanoma cell lines in vitro and in vivo. After transfection with liposomes containing the HuIFN-beta gene, human melanoma cell lines produced HuIFNbeta in the culture medium at levels ranging from 67 to 3.8 IU/ml on day 6, and growth of the cells was inhibited by 71-92%. Moreover, six injections of liposomes containing the HuIFNbeta gene completely eradicated human melanoma nodules transplanted onto the backs of nude mice 40 days after the first injection. Histological analysis of the injected nodules revealed that the HuIFNbeta gene transfection induced apoptosis of the human melanoma cells. These data suggest that transfection of the HuIFNbeta gene using cationic liposomes is a promising candidate for gene therapy of human melanoma.     

恶性黑色素瘤化疗研究进展

恶性黑色素瘤(Malignant melanoma,MM)是一种恶性程度极高的实体肿瘤,它极具侵袭性,预后差。尽管晚期黑色素瘤的治疗已经进入靶向和免疫治疗的时代,但是化疗仍然不可摒弃。恶性黑色素瘤的化疗经历了从单药化疗、两药或三药甚至四药的联合化疗、再到生物化疗这一发展过程。本文综述恶性黑色素瘤化疗研究进展,对主要的化疗方案进行比较,展望化疗未来发展的方向。     

Immune status of patients with malignant melanoma during combined therapy including local ultra-high frequency hyperthermia

Immune status was evaluated in patients with clinical stage I malignant melanoma randomized to receive either preoperative irradiation (27 Gy) alone or in combination with local microwave hyperthermia (before or after it). Mononuclear, total T-lymphocyte, T-helper, T-suppressor, B-lymphocyte and blood-immune complex levels were assayed. The analysis of immunologic parameters dynamics pointed to beneficial changes occurring in immunoregulating T-lymphocytes in patients receiving hyperthermia as a component of combined treatment.     

Adjuvant therapy for malignant melanoma

Malignant melanoma is increasing in incidence worldwide, and many patients remain at a significant risk of recurrence following surgical resection. Over the past 30 years, interferon-α has been the only agent approved for adjuvant therapy of melanoma. This review summarizes the rationale for adjuvant therapy, and discusses the roles of interferon, immunotherapy, chemotherapy and radiation therapy in the adjuvant setting. New approaches and novel combinations that appear promising for the adjuvant therapy of malignant melanoma are also outlined.     

恶性黑色素瘤的靶向治疗

目前恶性黑色素瘤(MM)主要的靶向治疗方法包括信号传导通路抑制剂、靶向凋亡、蛋白酶体抑制剂、抗血管生成治疗等.MM靶向治疗研究的发展方向是根据患者对靶向治疗敏感性的差异选择更加个体化的靶向治疗方法和技术.     

Targeted therapy for malignant melanoma

There has been much development in the field of targeted therapy for melanoma stemming from efforts to decrease treatment-related toxicities and enhance specific cytotoxicity. This review focuses on three modalities of targeted melanoma therapy based on the biology of the targeting mechanism. The first of these modalities is immunotherapy, which functions to generate a specific antimelanoma immunity. A second modality utilizes metabolic pathways of melanin synthesis to target melanoma cells specifically. A third modality ensues from recent advances in molecular biology and the identification of genes responsible for the malignant transformation of normal melanocytes to melanomas. This work has furthered our understanding of the basis of malignancy, as well as the development of novel strategies aimed at targeting aberrant growth in melanoma.     

Adjuvant therapy for malignant melanoma

Malignant melanoma is increasing in incidence worldwide, and many patients remain at a significant risk of recurrence following surgical resection. Over the past 30 years, interferon-alpha has been the only agent approved for adjuvant therapy of melanoma. This review summarizes the rationale for adjuvant therapy, and discusses the roles of interferon, immunotherapy, chemotherapy and radiation therapy in the adjuvant setting. New approaches and novel combinations that appear promising for the adjuvant therapy of malignant melanoma are also outlined.     

A case of unresectable gallbladder cancer responding to combination therapy with hyperthermia and local chemotherapy

A 78-year-old woman was admitted to our hospital for the control of gallbladder cancer. A peritoneal metastasis, diagnosed as unresectable cancer, was detected during surgery in a previous hospital, and a biliary stent was introduced and gastrojejunostomy was performed. In our hospital she was treated weekly with local chemotherapy (PFL; cisplatin 2.5-5 mg/body ia, fluorouracil 300 mg/body ia, and calcium folinate 30 mg/body ia, via the common hepatic arterial port) at the time of hyperthermia. Hyperthermia was performed with a Thermox 500 (HEH-500 C) at the power of 500 watts for 45-60 minutes. To enhance the hyperthermia effect, mitomycin C 2-4 mg/body ia via the common hepatic arterial port and 500 ml of 7.5% glucose infusion were given. As a result of the combination therapy, the volume of the whole tumor was reduced to 60.9% on computed tomography, and diagnosed as PR. The serum level of CA19-9 decreased from 3,000 U/ml to 300 U/ml. The patient continued to receive the therapy for 1 year, and is now well. Therefore, we conclude that combination therapy with hyperthermia and local chemotherapy seems beneficial in managing unresectable advanced gallbladder cancer, especially for the elderly.     

腹腔内联合灌注化疗治疗恶性腹膜间皮瘤的临床研究

目的探讨腹腔内联合化疗治疗恶性腹膜间皮瘤(malignant peritoneal mes-othelioma,MPM)的疗效。方法采用多柔比星30mg/m2+顺铂25mg/m2+丝裂霉素4～6mg联合化疗方案对19例MPM患者进行腹腔内注射,每周1次,每例共注射3～4次,分析腹水变化情况。结果近期疗效CR10例(52.6%),PR6例(31.6%),NC3例(15.8%),有效率(CR+PR)为84.2%,显著高于对照组(41.7%),χ2=6.09187,P<0.025。不良反应轻微。结论腹腔内联合化疗治疗MPM近期疗效好,不良反应少,是MPM的有效治疗方法之一。     

Adjuvant therapy for patients with high-risk malignant melanoma

The role of adjuvant therapy in the treatment of patients with high-risk malignant melanoma remains an area of intense investigation. The initial enthusiasm for high-dose interferon has been tempered by the results of more recent studies that allow for conflicting interpretations. Vaccine therapy trials have failed to clearly demonstrate a survival benefit, although several trials are currently ongoing. Recent studies of the role of chemotherapy suggest there may be combinations that have a survival benefit which deserve further study. This article will address patient selection and staging workup, and review options for treatment.     

尼妥珠单抗联合化疗治疗恶性胶质瘤

目的 评价尼妥珠单抗联合化疗治疗恶性胶质瘤的疗效及不良反应.方法 尼妥珠单抗200 mg/次,每周1次,连续8周后改为每2周1次;根据患者O6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)蛋白表达状况和既往化疗效果,采用个体化的化疗方案.结果 14例恶性胶质瘤患者共接受尼妥珠单抗治疗122次,中位治疗7.5次(2～20次).联合的化疗方案中,替莫唑胺21 d方案10例,替莫唑胺5 d力案2例,替尼泊甙联合顺铂方案1例,替尼泊甙联合尼莫司汀方案1例.PR 3例(21.4%),SD 6例(42.9%),客观有效率为21.4%,疾病控制率(PR+SD)为64.3%.中位无进展生存期(PFS)为4个月(95%CI0.7～7.3),6个月的疾病无进展生存率为30.6%.主要的不良反应为Ⅰ～Ⅱ度的中性粒细胞下降(2例)、血小板下降(2例)、淋巴细胞下降(1例)、恶心呕吐(3例)和无症状的转氨,升高(1例).1例替尼泊甙联合顺铂方案化疗的患者发生Ⅳ度中性粒细胞下降和血小板下降.1例患者出现尼妥珠单抗治疗相关痤疮样皮疹.结论 尼妥珠单抗联合化疗治疗恶性胶质瘤有一定疗效,患者耐受性好,值得进一步扩大病例数开展临床研究.

Abstract：

Objective Nimotuzumab is a humanized monoclonal antibody targeted against epidermal growth factor receptor (EGFR). Recent clinical studies show that patients with malignant gliomas could benefit from nimotuzumab treatment. The aim of the present study was to evaluate the efficacy and side effects of nimotuzumab in combination with chemotherapy for patients with malignant gliomas. Methods The patients received 200 mg of nimotuzumab infusion intravenously over 60 minutes once weekly for the first eight weeks and then once every two weeks until unacceptable toxicity or tumor progression occurred.Individualized chemotherapy was administered based on O6-methylguanine-DNA methyltransferase (MGMT)expression and previous chemotherapy responses in combined with nimotuzumab. Results Fourteen patients received a total of 122 times of nimotuzumab ranging from 2 to 20 ( median 7.5 times ). Combined chemotherapy regimens included:continuous 21-day temozolomide ( 10 cases), standard 5-day temozolomide (2 cases), teniposide plus cisplatin ( 1 case), and teniposide plus nimustine ( 1 case). Partial response (PR) and stable disease (SD) were found in 3 patients (21.4%)and 6 patients (42.9%), respectively.Disease control rate ( PR + SD) was 64.3%. The median progression-free survival (PFS) was 4 months (95%CI:0.7-7.3) and PFS at 6 months was 30. 6%. The most common toxicities include grade Ⅰ -Ⅱ neutropenia (2 cases), thrombocytopenia ( 2 cases), lymphopenia ( 1 case), nausea and vomitting ( 3case) and asymptomatic transaminase increase ( 1 case). One patient developed grade Ⅳ neutropenia and thrombocytopenia. One patient developed nimotuzumab-related acneiform rash. Conclusions Nimotuzumab in combination with chemotherapy has moderate activity in patients with malignant gliomas and the toxicities are well tolerable, therefore, worth further investigation.     

Combined treatment with radiation and hyperthermia in metastatic malignant melanoma

In 24 patients with metastatic malignant melanoma, combined treatment with radiation and hyperthermia was administered to 38 localizations, radiation alone to 8 comparative localizations and hyperthermia alone to 3 localizations. Hyperthermia was administered during one hour by using a 433 MHz microwave generator. The heat treatment was given within 30 min following irradiation. Although an intratumoral temperature of 43 degrees C was aimed, considerable variations occurred during one session and from session-to-session. Radiation schedules consisted in either one large fraction (6-8 Gy) once a week in 14-21 days or two fractions (4-5 Gy) twice a week in 21 days. In the group of patients receiving irradiation once a week, three heat treatments were administered. In the twice-a-week radiation schedule, six heat sessions were given. The overall complete response (CR) rate in patients receiving combined treatment was 50%. In the group of patients treated with hyperthermia and irradiation schedules of 8 Gy per fraction, the CR rate was 83%. Irradiation alone achieved 38% CR rate but some of these CR relapsed during follow-up whereas the comparative area treated with radiation and heat remained under control at this time. The lesions treated with heat alone did not show any response to treatment. Enhancement of the acute skin reactions was generally observed. However, because the total doses were relatively low, this enhancement did not constitute a clinical problem. CR appears to occur more frequently in small tumor sizes. The highest and lowest temperature ever registered during any session of hyperthermia did not seem to correlate with the tumor response.     

Cationic albumin-conjugated magnetite nanoparticles,novel candidate for hyperthermia cancer therapy

Abstract

We developed a novel hyperthermia material for cancer therapy, cationic albumin-conjugated magnetite nanoparticles (MNPs), which absorb the energy of an alternating magnetic field and convert it into heat. MNPs of about 10?nm were synthesised through co-precipitation, and citric acid was used to stabilise the MNP suspension. Then albumin was cationised by replacing anionic side chain groups with cationic groups. The surface modification of the MNPs was provided by cationic albumin, which was covalently conjugated to carboxylic acid functions located at the distal end of the MNPs’ surface by carbodiimide chemistry. Finally, we obtained stable superparamagnetic suspensions with particle sizes of 140?nm and saturation magnetisation of 67?emu/g, which do not have the disadvantage of eventual desorption of physical attachment. We also analysed the potential of these particles for magnetic fluid hyperthermia by determination of the specific absorption rate at a constant frequency of 215 kHz; the temperature increase of the particles was 30.8?°C. This study experimentally demonstrates the high efficiency of these nano-heaters.     

Effective combination chemo/hormonal therapy for malignant melanoma: experience with three consecutive trials.

Our experience with the combination of dacarbazine, carmustine, cisplatin with and without tamoxifen is reported. In our initial study, with all 4 drugs, we had an overall response rate of 50% with a complete response rate of 15%. Due to a high incidence of deep venous thrombosis and the lack of effectiveness of tamoxifen as a single agent, we deleted tamoxifen from the regimen and treated another 20 patients. Surprisingly, the response rate decreased to 10%. We then re-incorporated tamoxifen into the regimen and treated 25 additional patients. In this third group of patients we experienced an objective response rate of 52% with a complete response rate of 8%. Overall, 65 patients have been treated: 45 with and 20 without tamoxifen. Twenty-three (51%) patients treated with tamoxifen have responded, with 5 (11%) patients achieving a complete response. Only 2 (10%) patients treated without tamoxifen have responded. Despite the improvement in the response rate, a corresponding increase in survival has not been seen. Patients treated with tamoxifen had a mean survival of 10.8 (SD 13.6) months compared with a mean survival of 9.8 (SD 7.3) months for those treated without tamoxifen. The absence of survival advantage for the tamoxifen-treated patients may be due to early failure in the central nervous system. In 48% of the responding tamoxifen-treated patients, the first site of failure was the central nervous system, while systemic disease was still responding.