Mismatch repair gene germline mutations in patients with prostate cancer / 浙江大学学报·医学版

OBJECTIVES@#To investigate the prevalence of pathogenic germline mutations of mismatch repair (MMR) genes in prostate cancer patients and its relationship with clinicopathological characteristics.@*METHODS@#Germline sequencing data of 855 prostate cancer patients admitted in Fudan University Shanghai Cancer Center from 2018 to 2022 were retrospectively analyzed. The pathogenicity of mutations was assessed according to the American College of Medical Genetics and Genomics (ACMG) standard guideline, Clinvar and Intervar databases. The clinicopathological characteristics and responses to castration treatment were compared among patients with MMR gene mutation (MMR+ group), patients with DNA damage repair (DDR) gene germline pathogenic mutation without MMR gene (DDR+MMR－ group) and patients without DDR gene germline pathogenic mutation (DDR－ group).@*RESULTS@#Thirteen (1.52%) MMR+ patients were identified in 855 prostate cancer patients, including 1 case with MLH1 gene mutation, 6 cases with MSH2 gene mutation, 4 cases with MSH6 gene mutation and 2 cases with PMS2 gene mutation. 105 (11.9%) patients were identified as DDR gene positive (except MMR gene), and 737 (86.2%) patients were DDR gene negative. Compared with DDR－ group, MMR+ group had lower age of onset (P<0.05) and initial prostate-specific antigen (PSA) (P<0.01), while no significant differences were found between the two groups in Gleason score and TMN staging (both P>0.05). The median time to castration resistance was 8 months (95%CI: 6 months-not achieved), 16 months (95%CI: 12-32 months) and 24 months (95%CI: 21-27 months) for MMR+ group, DDR+MMR－ group and DDR－ group, respectively. The time to castration resistance in MMR+ group was significantly shorter than that in DDR+MMR－ group and DDR－ group (both P<0.01), while there was no significant difference between DDR+MMR－ group and DDR－ group (P>0.05).@*CONCLUSIONS@#MMR gene mutation testing is recommended for prostate cancer patients with early onset, low initial PSA, metastasis or early resistance to castration therapy.

Improving adenoviral vectors and strategies for prostate cancer gene therapy

Gene therapy has been evaluated for the treatment of prostate cancer and includes the application of adenoviral vectors encoding a suicide gene or oncolytic adenoviruses that may be armed with a functional transgene. In parallel, versions of adenoviral vector expressing the p53 gene (Ad-p53) have been tested as treatments for head and neck squamous cell carcinoma and non-small cell lung cancer. Although Ad-p53 gene therapy has yielded some interesting results when applied to prostate cancer, it has not been widely explored, perhaps due to current limitations of the approach. To achieve better functionality, improvements in the gene transfer system and the therapeutic regimen may be required. We have developed adenoviral vectors whose transgene expression is controlled by a p53-responsive promoter, which creates a positive feedback mechanism when used to drive the expression of p53. Together with improvements that permit efficient transduction, this new approach was more effective than the use of traditional versions of Ad-p53 in killing prostate cancer cell lines and inhibiting tumor progression. Even so, gene therapy is not expected to replace traditional chemotherapy but should complement the standard of care. In fact, chemotherapy has been shown to assist in viral transduction and transgene expression. The cooperation between gene therapy and chemotherapy is expected to effectively kill tumor cells while permitting the use of reduced chemotherapy drug concentrations and, thus, lowering side effects. Therefore, the combination of gene therapy and chemotherapy may prove essential for the success of both approaches.

A systematic review of the nutritional adequacy of the diet in the Central Andes / Revisión sistemática de la adecuación nutricional del régimen alimentario en los Andes centrales

OBJECTIVE:To examine dietary adequacy in the Andean area, including macro- and micronutrient intakes, with a particular focus on rural communities; to highlight nutrition priorities in the Andes; and to identify opportunities for improvement. METHODS: A comprehensive literature search was conducted, identifying published and grey literature in English and Spanish related to diet in the central Andean countries of Bolivia, Colombia, Ecuador, and Peru. Articles reporting data from dietary surveys or nutrition interventions were included. Thirty-four papers or reports published in 1969-2011 were included in the final review. The mean and variation in intakes by sex and age group of all presented nutrients were collated and the mean of means were calculated. RESULTS: Thiamin, niacin, and vitamin C intakes were usually adequate. Intakes of most other micronutrients, including iron, zinc, vitamin A, riboflavin, vitamin B12, folate, and zinc were low, likely resulting in high levels of inadequacy. Energy intakes were lower than requirements, but it is unlikely to be a common problem, rather, this result was probably due to the known tendency of most dietary survey tools to underreport intake. However, energy from fat intakes was very low, usually less than 20% of the total, and in some settings, less than 10%. CONCLUSIONS: The inadequate intake of some micronutrients is common in many developing countries, but the extremely low intake of dietary fat found in the central Andes is not. Increased consumption of animal-source foods would increase fat intakes, while addressing micronutrient deficiencies; however, the impact on the fragile ecosystem of the Andes needs considering. Indigenous crops, such as lupine bean, quinoa, and amaranth are also rich in fat or micronutrients.

OBJETIVO: Analizar la adecuación del régimen alimentario en la zona andina, incluidas las ingestas de macro y micronutrientes, prestando especial atención a las comunidades rurales; señalar las prioridades nutricionales en los Andes; y establecer las oportunidades de mejora. MÉTODOS: Se llevó a cabo una exhaustiva búsqueda bibliográfica, en la que se seleccionaron documentos publicados y procedentes de la literatura gris, en inglés y español, relacionados con el régimen alimentario en los países andinos centrales de Bolivia, Colombia, Ecuador y Perú. Se incluyeron artículos que aportaran datos de encuestas alimentarias o intervenciones nutricionales. En el análisis final, se incluyeron 34 artículos o informes publicados desde 1969 a 2011. Se recopilaron las medias y las variaciones de las ingestas de todos los nutrientes presentados según el sexo y el grupo de edad, y se calculó la correspondiente media de las medias. RESULTADOS: Las ingestas de tiamina, niacina y vitamina C eran generalmente adecuadas. Las ingestas de la mayor parte de los restantes micronutrientes, incluidos el hierro, el cinc, la vitamina A, la riboflavina, la vitamina B12 y el folato, eran bajas, lo que probablemente ocasionaba altos niveles de inadecuación. Los aportes energéticos eran inferiores a los requeridos, aunque es poco probable que ello constituya un problema frecuente; más bien, este resultado podría deberse a la tendencia conocida de notificar insuficientemente la ingesta en la mayor parte de las encuestas alimentarias. Sin embargo, el aporte energético procedente del consumo de grasas era muy reducido, generalmente por debajo del 20% del total, y en algunos lugares, por debajo del 10%. CONCLUSIONES: La ingesta inadecuada de algunos micronutrientes es frecuente en muchos países en desarrollo, aunque no es tan frecuente la ingesta extremadamente baja de grasa alimentaria observada en los Andes centrales. Un mayor consumo de alimentos de origen animal aumentaría la ingesta de grasas, al tiempo que abordaría las carencias en micronutrientes; sin embargo, debe tenerse en cuenta su posible repercusión sobre el frágil ecosistema de los Andes. Los cultivos autóctonos, como el frijol de altramuz, la quinoa y el amaranto, son también ricos en grasas o micronutrientes.

Controversies in using urine samples for prostate cancer detection: PSA and PCA3 expression analysis

PURPOSE: Prostate cancer (PCa) is one of the most commonly diagnosed malignancies in the world. Although PSA utilization as a serum marker has improved prostate cancer detection it still presents some limitations, mainly regarding its specificity. The expression of this marker, along with the detection of PCA3 mRNA in urine samples, has been suggested as a new approach for PCa detection. The goal of this work was to evaluate the efficacy of the urinary detection of PCA3 mRNA and PSA mRNA without performing the somewhat embarrassing prostate massage. It was also intended to optimize and implement a methodological protocol for this kind of sampling. MATERIALS AND METHODS: Urine samples from 57 patients with suspected prostate disease were collected, without undergoing prostate massage. Increased serum PSA levels were confirmed by medical records review. RNA was extracted by different methods and a preamplification step was included in order to improve gene detection by Real-Time PCR. RESULTS: An increase in RNA concentration with the use of TriPure Isolation Reagent. Despite this optimization, only 15.8 percent of the cases showed expression of PSA mRNA and only 3.8 percent of prostate cancer patients presented detectable levels of PCA3 mRNA. The use of a preamplification step revealed no improvement in the results obtained. CONCLUSION: This work confirms that prostate massage is important before urine collection for gene expression analysis. Since PSA and PCA3 are prostate specific, it is necessary to promote the passage of cells from prostate to urinary tract, in order to detect these genetic markers in urine samples.

Construction of a recombinant adenovirus co-expressing truncated human prostate-specific membrane antigen and mouse 4-1BBL genes and its effect on dendritic cells

Our aim was to construct a recombinant adenovirus co-expressing truncated human prostate-specific membrane antigen (tPSMA) and mouse 4-1BBL genes and to determine its effect on dendritic cells (DCs) generated from bone marrow suspensions harvested from C57BL/6 mice for which the effect of 4-1BBL on DCs is not clear, especially during DCs processing tumor-associated antigen. Replication deficient adenovirus AdMaxTM Expression System was used to construct recombinant adenovirus Ad-tPSMA-internal ribosome entry site-mouse 4-1BBL (Ad-tPSMA-IRES-m4-1BBL) and Ad-enhanced green fluorescent protein. Day 7 proliferating DC aggregates generated from C57BL/6 mice were collected as immature DCs and further mature DCs were obtained by lipopolysaccharide activated immature DCs. After DCs were exposed to the recombinant adenovirus with 250 multiplicity of infection, the expression of tPSMA and m4-1BBL proteins were detected by Western blot, and the apoptosis and phenotype of DCs were analyzed by flow cytometry. Cytokines (IL-6 and IL-12) in the supernatant were detected by enzyme-linked immunosorbent assay (ELISA). Proliferation of T cells was detected by allogeneic mixed lymphocyte reactions. The tPSMA and m4-1BBL proteins were expressed correctly. The apoptosis rate of DCs transfected with Ad-tPSMA-IRES-m4-1BBL was 14.6 percent, lower than that of control DCs. The expression of co-stimulatory molecules [CD80 (81.6 ± 5.4 percent) and CD86 (80.13 ± 2.81 percent)] up-regulated in Ad-tPSMA-IRES-m4-1BBL-pulsed DCs, and the level of IL-6 (3960.2 ± 50.54 pg/mL) and IL-12 (249.57 ± 12.51 pg/mL) production in Ad-tPSMA-IRES-m4-1BBL-transduced DCs were significantly higher (P < 0.05) than those in control DCs. Ad-tPSMA-IRES-m4-1BBL induced higher T-cell proliferation (OD450 = 0.614 ± 0.018), indicating that this recombinant adenovirus can effectively enhance the activity of DCs.