Radioiodine therapy for castration-resistant prostate cancer following prostate-specific membrane antigen promoter-mediated transfer of the human sodium iodide symporter

Radioiodine therapy, the most effective form of systemic radiotherapy available, is currently useful only for thyroid cancer because of the thyroid-specific expression of the human sodium iodide symporter （hNIS）. Here, we explore the efficacy of a novel form of gene therapy using prostate-specific membrane antigen （PSMA） promoter-mediated hNIS gene transfer followed by radioiodine administration for the treatment of castration-resistant prostate cancer （CRPC）. The androgen-dependent C33 LNCaP cell line and the androgen-independent C81 LNCaP cell line were transfected by adenovirus. PSMA promoter-hNIS （Ad.PSMApro-hNIS） or adenovirus.cytomegalovirus-hNIS containing the cytomegalovirus promoter （Ad.CMM-hNIS） or a control virus. The iodide uptake was measured in vitro. The in vivo iodide uptake by C81 cell xenografts in nude mice injected with an adenovirus carrying the hNIS gene linked to PSMA and the corresponding tumor volume fluctuation were assessed. Iodide accumulation was shown in different LNCaP cell lines after Ad.PSMApro-hNIS and Ad.CMV-hNIS infection, but not in different LNCaP cell lines after adenovirus.cytomegalovirus （Ad.CMV） infection. At each time point, higher iodide uptake was shown in the C81 cells infected with Ad.PSMApro-hNIS than in the C33 cells （P 〈 0.05）. An in vivo animal model showed a significant difference in 1311 radioiodine uptake in the tumors infected with Ad.PSMApro-hNIS, Ad.CMV-hNIS and control virus （P 〈 0.05） and a maximum reduction of tumor volume in mice infected with Ad.PSMApro-hNIS. These results show prostate-specific expression of the hNIS gene delivered by the PSMA promoter and effective radioiodine therapy of CRPC by the PSMA promoter-driven hNIS transfection.

A biomimetic antitumor nanovaccine based on biocompatible calcium pyrophosphate and tumor cell membrane antigens

Currently,the cancer immunotherapy has made great progress while antitumor vaccine attracts substantial attention.Still,the selection of adjuvants as well as antigens are always the most crucial issues for better vaccination.In this study,we proposed a biomimetic antitumor nanovaccine based on biocompatible nanocarriers and tumor cell membrane antigens.Briefly,endogenous calcium pyrophosphate nanogranules with possible immune potentiating effect are designed and engineered,both as delivery vehicles and adjuvants.Then,these nanocarriers are coated with lipids and B16-OVA tumor cell membranes,so the biomembrane proteins can serve as tumor-specific antigens.It was found that calcium pyrophosphate nanogranules themselves were compatible and possessed adjuvant effect,while membrane proteins including tumor associated antigen were transferred onto the nanocarriers.It was demonstrated that such a biomimetic nanovaccine could be well endocytosed by dendritic cells,promote their maturation and antigen-presentation,facilitate lymph retention,and trigger obvious immune response.It was confirmed that the biomimetic vaccine could induce strong T-cell response,exhibit excellent tumor therapy and prophylactic effects,and simultaneously possess nice biocompatibility.In general,the present investigation might provide insights for the further design and application of antitumor vaccines.

Prostate-specific membrane antigen expression in hepatocellular carcinoma,cholangiocarcinoma,and liver cirrhosis

BACKGROUND Primary liver cancer includes three subtypes:Hepatocellular carcinoma(HCC),intrahepatic cholangiocarcinoma(CCA),and combined hepatocellular carcinoma.Patients with primary liver cancer experienced poor prognosis and high mortality,so early detection of liver cancer and improved management of metastases are both key strategies to reduce the death toll from liver cancer.Prostate-specific membrane antigen(PSMA)expression in the tumor-associated neovasculature of nonprostate malignancies including liver cancer has been reported recently,but conclusive evidence of PSMA expression based on the pathological type of liver cancer remains limited.AIM To study the expression of PSMA in HCC,CCA,and liver cirrhosis.METHODS A total of 446 formalin-fixed paraffin-embedded(FFPE)liver tumor and liver cirrhosis tissue samples were obtained retrospectively from the Pathology Department of Tongji Hospital.Immunohistochemistry was used to detect PSMA expression in these 446 FFPE liver biopsy specimens(213 HCC,203 CCA,and 30 liver cirrhosis).The tumor compartment and the associated neovascular endothelium were separately analyzed.PSMA expression was examined by two certified pathologists,and the final results were presented in a 4-point scoring system(0-3 points).Correlation between PSMA expression and clinicopathological information was also assessed.RESULTS PSMA was expressed primarily in the neovascular endothelium associated with tumors.The positive rate of PSMA staining in HCC was significantly higher than that in CCA(86.8%vs 79.3%;P=0.001)but was only 6.6%in liver cirrhosis(P=0.000).HCC cases had more 3-score PSMA staining than CCA had(89/213,41.8%vs 35/203,17.2%;P=0.001).PSMA expression correlated positively with the stage and grade of HCC and CCA.In both liver cancer subtypes,there were more PSMA+cases in stages III–V diseases than in stages I and II.High staining intensity of PSMA was more frequently observed in liver cancers at high grade and advanced stage.There was no significant association of PSMA expression with sex,age,region,α-fetoprotein,hepatitis B surface antigen,or tumor size in both tumor subtypes.CONCLUSION Neovascular PSMA may be a promising marker to differentiate HCC from liver cirrhosis and a prognostic marker for anti-tumor angiogenesis therapy for HCC.

Determination of the genus-specific antigens in outer membrane proteins from the strains of Leptospira interrogans and Leptospira biflexa with different virulence

Objective:To determine the existence of genus-specific antigens in outer membrane proteins (OMPs) of leptospira with different virulence. Methods: Microscope agglutination test (MAT) was applied to detect the agglutination between commercial rabbit antiserum against leptospiral genus-specific TR/Patoc I antigen and 17 strains of Leptospira interrongans belonging to 15 serogroups and 2 strains of Leptospira biflexa belonging to 2 serogroups.The outer envelopes (OEs) of L.interrogans serogroup Icterohaemorrhagiae serovar lai strain lai (56601) with strong virulence and serogroup Pomona serovar pomona strain Luo (56608) with low virulence,and L.biflexa serogroup Semaranga serovar patoc strain Patoc I without virulence were prepared by using the method reported in Auran et al.(1972).OMPs in the OEs were obtained by treatment with sodium deoxycholate. SDS-PAGE and western blot were used for analyzing the features of the OMPs on electrophoretic pattern and the immunoreactivity to the antiserum against TR/Patoc I antigen, respectively. Results:All the tested strains belonging to different leptospiral serogroups agglutinated to the antiserum against leptospiral genus-specific TR/Patoc I antigen with agglutination titers ranging from 1:256-1:512. A similar SDS-PAGE pattern of the OMPs from the three strains of leptospira with different virulence was shown and the molecular weight of a major protein fragment in the OMPs was found to be approximately 60 KDa.A positive protein fragment with approximately 32 KDa confirmed by Western blot,was able to react with the antiserum against leptospiral genus-specific TR/Patoc I antigen, and was found in each the OMPs of the three stains of leptospira.Conclusion: There are genus-specific antigens on the surface of L.interrogans and L.biflexa. The OMP with molecular weight of 32 KDa may be one of the genus-specific protein antigens of leptospira.

Immunogenicity and protective role of antigenic regions from five outer membrane proteins of Flavobacterium columnare in grass carp Ctenopharyngodon idella

Flavobacterium columnare causes columnaris disease in freshwater fi sh. In the present study, the antigenic regions of fi ve outer membrane proteins(OMPs), including zinc metalloprotease, prolyl oligopeptidase, thermolysin, collagenase and chondroitin AC lyase, were bioinformatically analyzed, fused together, and then expressed as a recombinant fusion protein in Escherichia coli. The expressed protein of 95.6 k Da, as estimated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was consistent with the molecular weight deduced from the amino acid sequence. The purifi ed recombinant protein was used to vaccinate the grass carp, C tenopharyngodon idella. Following vaccination of the fi sh their Ig M antibody levels were examined, as was the expression of I g M, Ig D and Ig Z immunoglobulin genes and other genes such as MHC Iα and MHC I I β, which are also involved in adaptive immunity. Interleukin genes( IL), including I L- 1β, IL- 8 and I L- 10, and type I and type II interferon(I FN) genes were also examined. At 3 and 4 weeks post-vaccination(wpv), signifi cant increases in Ig M antibody levels were observed in the fi sh vaccinated with the recombinant fusion protein, and an increase in the expression levels of I g M, Ig D and Ig Z genes was also detected following the vaccinations, thus indicating that an adaptive immune response was induced by the vaccinations. Early increases in the expression levels of IL and IFN genes were also observed in the vaccinated fi sh. At four wpv, the fi sh were challenged with F. column a re, and the vaccinated fi sh showed a good level of protection against this pathogen, with 39% relative percent survival(RPS) compared with the control group. It can be concluded, therefore, that the fi ve OMPs, in the form of a recombinant fusion protein vaccine, induced an immune response in fi sh and protection against F. columnare.

Analysis of sperm membrane antigens relevant to antisperm antibody using Western blot

To identify the sperm membrane antigens associated with antisperm antibody. Methods: The antisperm antibody in serum was tested by ELISA. Antisperm antibody positive sera from 18 infertile men and 15 infertile women were used. The molecular weight (MW) of sperm membrane antigens associated with the antisperm antibody was analyzed with antisperm antibody positive serum using Western blot. Results: Eight kinds of MW of sperm membrane antigens were identified. The ratio of identification on the 78 KD(60.7 %), 60KD (71.4 %), 51 KD (14.9 %) and 23 KD (14.29 %) sperm antigen was higher than others. Conclusion: Sperm membrane antigens with MW of 78 KD, 60 KD, 51 KD and 23 KD were associated with antisperm antibody and immunological infertility. (Chin J Andro12002; 16: 345)

The Expression of Sperm Membrane Peptide-Hepatitis B SurfaceAntigen Fusion Protein with Recombinant Vaccinia Virus

A synthetic oligonucleotide, HSD-2a, encoding a peptide segment of the extracel-lular domain of a human sperm membrane protein, YWK-II, was fused with hepatitisB surface antigen gene (HBs gene ). The fused gene was then cloned to pUC18plasmid. The fused gene was prepared from the recombinant pUC18 plasmid byBamH I and Eco R I digestion, and then cloned into the transfer vector pGJP- 5 underthe control of P;., promoter, designated as pGJP-HSD/HBs. CV-1 cells were co-transfected with vaccinia virus (Tian Tan strain) and pGJP-HSD/IIBs and homolo-gous re combination occurred between the vaccinia virus TK gene of the plasmid flank-ing the foreign gene and the same sequences within the virus genome. TK phen0tyPerecombinant virus, vv-HSD/HBs, were selected from trandected HuTK' cells byplaque purthcation technique. The eopressi0n of HSD-b in spent medium and cellu-lar protein of HuTK cells infected with vv-HSD/HBs was determined by ELISAand Western-blot analysis using anti-rwK-II antiserum. The present study indicatesthat the vv-HSD/HBs seems promising as an antdertility vaccine.

STRUCTURE AND EXPRESSION OF EPSTEIN-BARR VIRUS MEMBRANE ANTIGEN IN RECOMBINANT VACCINIA VIRUS

The Epstein-Barr virus membrane antigen was constructed and inserted into vaccinia virus, Tian-tan strain in order to study the effect of this virus on EB infection and tumorogenesis. The EBV-derived membrane antigen was expressed under the control of a 7.5 K promoter of vaccinia virus. The antibody against the membrane antigen of EB virus was produced on rabbits vaccinated with recombinant vaccinia virus.

GENE ENGINEERING EB VIRUS MEMBRANE ANTIGEN IN DETECTION OF MA-IgA ANTIBODY(COMPARISON WITH VCA-IgA AND EA-IgA ANTIBODIES)

With gene engineering EB virus membrane antigen as the diagnostic antigen, indirect immunofluo-rescence (IF) assay was used to detect IgA antibody against EB virus membrane antigen (MA-IgA) in sera from 202 nasopharyngeal carcinoma (NPC) patients and 315 controls (normal and patients with other tumors). MA-IgA antibody was positive in 96.8% of the pretreatment NPC patients with a GMT of 1:36.3. MA-IgA detection by this method was more sensitive than EA-IgA detection by IE. In contrast, patients with tumors other than NPC were negative for MA-IgA antibody. 9.1% of VCA-IgA positive persons were MA-IgA positive with a GMT of less than 1:5. No MA-IgA positive was found in VCA-IgA negatives. The results indicated that this method was relatively specific. In the treatment group, the positive rate and GMT of MA-IgA antibody declined with increase in survival time and the decline was faster than VCA-IgA. When recurrence or distant metastasis developed, similar to VCA-IgA and EA-IgA antibodies, the positive rate and GMT of MA-IgA antibody increased to its pretreatment level. Therefore, MA-IgA detection might be valuable in the early diagnosis and monitor of NPC.

New frontiers in focal therapy for prostate cancer:Prostate-specific membrane antigen positron emission tomography/magnetic resonance imaging

Imaging has a central role in the context of focal therapy(FT)for prostate cancer(PCa).Prostate-specific membrane antigen(PSMA)positron emission tomography/magnetic resonance imaging(PET/MRI)is a novel imaging modality that combines the morpho-functional information of MRI with the molecular characterization of PET.Some papers reported the potential advantages of PSMA PET/MRI in different clinical scenarios.Limited evidence on PSMA PET/MRI is available in the setting of FT.PSMA PET/MRI can be an effective imaging modality for detecting primary PCa and seems to provide accurate local staging of primary PCa.PSMA PET/MRI also shows high performance for restaging and detecting tumor recurrence.The higher soft-tissue contrast and the reduction of ionizing radiation are the main advantages reported in the literature compared to PET/computed tomography.PSMA PET/MRI could represent a turning point in the management of patients with PCa in the context of FT.Further studies are needed to confirm its applications in this specific clinical setting.

Anti-tumor effects induced by gene vaccines co-expressing truncated human prostate specific membrane antigen gene and mouse 4-1BBL

Objective To investigate the influence of m4-1BBL on anti-tumor effects induced by truncated human prostate specific membrane antigen ( tPSMA ) gene in mice. Methods A eukaryotic expression plasmid encoding tPSMA and m4-1BBL ( pDC316-tPSMA-IRES m4-1BBL) ,pDC316-tPSMA and pDC316 were constructed.

Membrane redistributions through multi-intercellular exchanges and serial trogocytosis

Trogocytosis is a rapid transfer between cells of membranes and associated proteins. Trogocytic exchanges have been investigated between different cell types, mainly in two-cell systems, involving one donor and one acceptor cell type. Here, we studied trogocytosis in a more complex system, involving not only several immune cell subsets but also multiple tumor cells. We show that CD4~ T cells, CD8＋ T cells and monocytes can acquire membrane patches and the intact proteins they contain from different tumor cells by multiple simultaneous trogocytoses. The trogocytic ca- pabilities of CD4~ and CD8~ T cells were found to be similar, but inferior to that of autologous monocytes. Activated peripheral-blood mononuclear cells （PBMCs） may also exchange membranes between themselves in an all-autolo- gous system. For this reason, monocytes are capable of acquiring membranes from multiple tumor cell sources, and transfer them again to autologous T cells, along with some of their own membranes （serial trogocytosis）. Our data illustrate the extent of membrane exchanges between autologous activated immune effector cells and their environ- ment, and how the cellular content of the local environment, including ＂bystander＂ cells, may impact the functions of immune effector cells.

Bacterial outer membrane vesicle-based cancer nanovaccines

Tumor vaccines,a type of personalized tumor immunotherapy,have developed rapidly in recent decades.These vaccines evoke tumor antigen-specific T cells to achieve immune recognition and killing of tumor cells.Because the immunogenicity of tumor antigens alone is insufficient,immune adjuvants and nanocarriers are often required to enhance anti-tumor immune responses.At present,vaccine carrier development often integrates nanocarriers and immune adjuvants.Among them,outer membrane vesicles(OMVs)are receiving increasing attention as a delivery platform for tumor vaccines.OMVs are natural nanovesicles derived from Gramnegative bacteria,which have adjuvant function because they contain pathogen associated molecular patterns.Importantly,OMVs can be functionally modified by genetic engineering of bacteria,thus laying a foundation for applications as a delivery platform for tumor nanovaccines.This review summarizes 5 aspects of recent progress in,and future development of,OMV-based tumor nanovaccines:strain selection,heterogeneity,tumor antigen loading,immunogenicity and safety,and mass production of OMVs.

A New Vaccine Strategy of Dendritic Cell Presented Kinetoplastid Membrane(KMP-11)as Immunogen for Control against Experimental Visceral Leishmaniasis

Available reports suggest that, Leishmania donovani antigen KMP-11 may be significant in the modulation of immune responses in visceral leishmaniasis (VL). This study evaluated vaccine prospect of presentation of KMP-11 antigen through murine dendritic cells against VL in infected BALB/c mice. We report here that immunization with KMP-11 delivered through bone marrow derived dendritic cells can lead to killing of L. donovani in infected BALB/c mice. Furthermore, the strategy to use KMP-11 as vaccine delivered through DCs can stimulate the production of IFN-g, IL-12, IL-2R and TNF-α with concomitant down-regulation of IL-10 and IL-4. Furthermore, anti-leishmanial defence function (ROS) of splenocytes was observed increased in the presence of DC-delivered KMP-11 vaccination accompanied with an increased p38-MAPK signalling in vaccinated splenocytes. We summarized from our data that KMP-11 delivered through DCs has potential for eliciting protective immunity through pro-inflammatory cytokines (IFN-γ, IL-12, IL-2, TNF-α) following an up-regulation in signalling event of p38-MAPK. Therefore the study suggests a new control strategy against VL in future.

^(161)Tb-PSMA放射性配体治疗前列腺癌研究进展

前列腺特异性膜抗原(PSMA)放射性配体治疗(PRLT)已成为一种重要的新型疗法,尤其适合治疗常规疗法无法有效控制的转移性去势抵抗性前列腺癌(mCRPC),以镥-177(^(177)Lu)-PSMA-617为代表的PRLT具有良好的安全性,能有效延长mCRPC患者生存期,改善生活质量等优势。尽管^(177)Lu-PSMA-617治疗mCRPC取得成功,但仍有部分患者不敏感,甚至对治疗无响应,出现复发情况。铽-161(^(161)Tb)与177Lu均具有β-射线,但与^(177)Lu相比,其最大的优势在于释放针对小病灶治疗的俄歇电子和内转化电子,这种β-射线与俄歇电子的协同疗法,具有满足不同大小病灶治疗的优点,使得全球范围对^(161)Tb进行肿瘤治疗的关注度正在上升。本文将从^(161)Tb的制备、标记、质控、辐射剂量、临床前研究、临床研究以及展望等方面对^(161)Tb-PSMA研究进行综述,旨在为开展PRLT的医务人员及患者提供参考。

新型前列腺癌显像剂^(68)Ga-PSMA-4PY的初步临床研究

目的:评估一种体内快速清除的前列腺癌显像剂^(68)Ga-PSMA-4PY在临床应用中的潜力。方法:采用前体药盒化方法标记^(68)Ga-PSMA-4PY,进行质量控制确保其纯度和安全性;通过动态正电子发射体层成像(positron emission tomography,PET)分析^(68)Ga-PSMA-4PY在小鼠血液、肝脏和肾脏中的放射性摄取及清除。并通过临床显像研究比较^(68)Ga-PSMA-4PY与^(68)Ga-PSMA-11在前列腺癌患者体内的分布特性。结果:经前体药盒化标记的^(68)Ga-PSMA-4PY具有良好的标记效率和稳定性。小鼠动态PET结果显示,^(68)Ga-PSMA-4PY在血液中的清除特征与^(68)Ga-PSMA-11相近;^(68)Ga-PSMA-4PY在肾脏中的放射性摄取峰值出现在2 min内,并随后快速排出,而^(68)Ga-PSMA-11在肾脏中呈持续性增加,直至60 min时达到^(68)Ga-PSMA-4PY的11倍。临床研究表明,^(68)Ga-PSMA-4PY可以在前列腺癌中浓集,在肝、肾和脾脏中的放射性滞留明显低于^(68)Ga-PSMA-11,且唾液腺摄取降低了57.5%。结论:^(68)Ga-PSMA-4PY能够快速浓聚到PSMA阳性的肿瘤中,并迅速从正常组织中清除,将有助于减少非特异性摄取引起的不良反应,是一种具有良好前景的前列腺癌显像剂。

^(177)Lu-前列腺特异性膜抗原放射性配体疗法治疗前列腺癌临床应用专家共识

^(177)Lu标记的前列腺特异性膜抗原(prostate specific membrane antigen,PSMA)放射性配体疗法已在国外获批应用于晚期前列腺癌治疗,且正在国内开展多项临床试验。专家组参考国外经验和观点,并结合国内临床实践和实测数据,形成^(177)Lu-PSMA放射性配体疗法在前列腺癌临床应用中的专家共识。

靶向PSMA诊疗一体化:应用与进展

前列腺特异性膜抗原(prostate-specific membrane antigen,PSMA)最早在前列腺癌细胞中发现,是前列腺癌等高表达PSMA肿瘤诊疗的重要靶点。多种显像和治疗核素标记的PSMA不同配体在前列腺癌诊疗一体化中显示出重要作用。本文就靶向PSMA诊疗一体化的相关研究和临床应用进展进行述评。

PSMA PET/CT在局限性或局部进展性前列腺癌诊疗中的应用

前列腺特异性膜抗原正电子发射计算机断层扫描(PSMA PET/CT)是新兴的成像技术,近年来逐渐被应用于局限性或局部进展性前列腺癌的诊断及治疗中。PSMA PET/CT检查可有效减少不必要的活检且其引导下的靶向穿刺活检可显著提高临床显著性前列腺癌(csPCa)的检出率。基于PSMA PET的多影像融合穿刺活检技术可能是未来前列腺癌诊断研究的重点,与传统影像学技术相比,PSMA PET/CT在检测淋巴结转移方面具有优越性。PSMA PET/CT不仅可用于预测Gleason评分、手术切缘情况、术后病理等级等病理特征,还可用于评估前列腺癌患者新辅助治疗疗效、指导制定放射治疗方案、预测生化复发以及检出复发病灶。目前仍需进行更多大规模前瞻性研究进一步证实PSMA PET/CT在局限性或局部进展性前列腺癌诊疗中的应用价值。

^(177)Lu-PSMA放射性配体疗法治疗前列腺癌的临床实践专家共识(2024年版)

镥-177标记前列腺特异性膜抗原放射性配体疗法(^(177)Lu-labeled prostate-specific membrane antigen radioligand therapy,^(177)Lu-PSMA-RLT)是一种通过精准地向前列腺癌细胞递送^(177)Lu释放的β射线,引发肿瘤细胞DNA的辐射损伤从而杀灭肿瘤的创新治疗方法。现阶段国内外多个前列腺癌领域权威指南与共识推荐其用于治疗转移性去势抵抗性前列腺癌(metastatic castrate-resistant prostate cancer,mCRPC),国内尚未见关于^(177)Lu-PSMA-RLT治疗前列腺癌的临床实践路径和用药规范的报道。本共识由中国抗癌协会肿瘤核医学专业委员会和中国医师协会核医学医师分会共同发起,结合现有的^(177)Lu-PSMA-RLT注册研究结果、真实世界数据及国内外放射性核素临床治疗经验,根据牛津循证医学中心临床证据推荐等级和证据级别来评价不同级别的证据,在适应证、患者筛选、用药不良反应、随访和辐射安全等方面给出推荐意见。本共识旨在为临床医师提供^(177)Lu-PSMA-RLT治疗前列腺癌的临床参考,为进一步制订行业相关指南奠定基础。