Precision Medicine of Cancer

癌症的精确医学是多方面的，我们的研究，从长凳到床边的方法，已经导致了重大发现，将促进生物标记物的诊断和预后利用，并阐明微生物组在肺癌发生中所起的作用和机制。此外，研究健康差距仍然是一个焦点(Graff，M.等人，Am J Hum Genet。108：564-582,2021；蒋明等人。肺癌152：58-65,2021年)，这是建立在实验室的遗产的基础上，已经是第一个在美国人口的非洲遗产(Zanetti Ka)进行GWA。等人，肺癌98：33-42,2016)。癌症微生物组：最近对微生物区系组成的一项研究显示，在几种曾经被认为是不育的癌症类型中存在丰富的细菌(GreaTower KL。等人，癌细胞38：158-160,2020)。我们在腺癌和鳞癌中都发现了与TP53突变和吸烟相关的嗜酸杆菌。利用小鼠肺癌模型，我们证明了嗜酸杆菌是肺癌发生的贡献者，而不是乘客，通过激活促炎症的肺中性粒细胞。我们的数据显示，与PBS(Sham)对照组小鼠相比，暴露于A.temans(Acidovorax Temans)的小鼠有明显更大的肿瘤负担和更丰富的肺部损伤。在荷瘤小鼠中，灌输金黄色葡萄球菌不利于生存，反复暴露会加速肺癌的发展，因此，金黄色葡萄球菌在肺癌的发生中具有一定的作用。全组织RNA测序(RNAseq)比较了灌输沙门氏菌的小鼠和假对照组的差异基因表达，结果显示，通过MyD88依赖的信号转导，对沙门氏菌产生了强烈的炎症反应，这是启动先天和获得性免疫的关键途径，并有助于癌症的进展。在与Romina Goldszmid的合作中，我们研究了滴注A.temans后肿瘤免疫微环境的变化。我们的荧光激活细胞分选(FACS)分析结果表明，在中性粒细胞、CD4+ve、CD8+ve、双阴性(DN)T细胞的驱动下，温带沙门氏菌感染的小鼠肺中CD45+ve免疫细胞数增加，γ-Delta T细胞数也增加。总体而言，我们的结果表明，金黄色葡萄球菌增加了促炎细胞的渗透，这种细胞随着时间的推移而持续，促进了肺腺癌的发展。嗜中性粒细胞在KPC小鼠肺部的募集在被A.temans攻击时加剧。浸润性中性粒细胞的表型增加了CD44低ICAM1高SiglecF+PDL1+中性粒细胞的频率，以前的特征是长寿、成熟的促肿瘤相关的中性粒细胞，对KPC小鼠的肺癌进展很重要。RNAseq显示，在金黄色葡萄球菌挑战的小鼠中，激活的和成熟的中性粒细胞的特征得分更高。细菌暴露可能是中性粒细胞成熟的原因，所以我们比较了以前报道的致病性大肠杆菌(E.Coli)感染的中性粒细胞标志的得分，发现由于细菌感染，灌输沙门氏菌的小鼠有更高的中性粒细胞成熟标志得分。中性粒细胞是肺癌患者肿瘤微环境中的主要免疫细胞群，有证据表明，循环中的中性粒细胞与白细胞的比率增加与预后差和对包括免疫治疗在内的治疗反应差有关。下一步是开发一种治疗性的小鼠模型，并验证中性粒细胞在调节沙门氏菌诱导的肺肿瘤发生中的作用。癌症代谢组：识别的代谢物与特定癌症的相关性创建了可用于许多类型人类癌症的非侵袭性诊断和预后评估的生物标记物图谱。尿、血清和血浆的液体活检用于检测四种生物标志物(肌酸核苷(CR)、N-乙酰纽米酸(NAA)、皮质醇硫酸盐(CS)和27α-或-5-β-胆烷-3α、7α、12α24α、25α戊醇葡萄糖醛酸苷(NCPG))。生物标志物Prev.2016年25：978-86)。与其他已识别的尿代谢物生物标志物(如NaNA)配合使用，可提高诊断能力和可靠性。这些基础性研究验证了尿代谢物筛查的使用，从而进一步研究了生物标记物与人类癌症的关系。(民主党帕特尔等人的研究。J·帕姆·比默德的肛门。在2020年的年度报告中提到，尿代谢物生物标志物分析可以为肝内胆管细胞癌(ICC)提供诊断和预后评估。使用UPLC-MS/MS，四种代谢物，用于定量代谢物CR，N-乙酰神经氨酸(NANA)，皮质醇硫酸盐，和一个被命名为561+的葡萄糖醛酸碎片离子，在肝癌和ICC中显著增加，并与临床使用的标志物CA19-9相结合，在ICC分类方面具有很强的可靠性。NCI-MD队列进行了研究，观察结果得到了Tiger-LC队列的验证。我们确定了对生物标志物具有重要意义的属性，以用于液体活检中基于CLIA的生物标志物分析。

Precision medicine in cancer is multifaceted and our research, following the bench to bedside approach, has led to significant discoveries that will advance the diagnostic and prognostic utilization of biomarkers and elucidate the role and mechanism the microbiome plays in lung carcinogenesis. In addition, researching health disparity remains a focus (Graff, M. et al., Am J Hum Genet. 108: 564-582, 2021; Jiang M. et al. Lung Cancer 152: 58-65, 2021) that build on the laboratory's legacy of having been the first to conduct GWAS in the US population of African heritage (Zanetti KA. et al., Lung Cancer 98: 33-42, 2016). Cancer Microbiome: A recent study of microbiota composition revealed an abundance of bacteria in several cancer types once thought to be sterile (Greathouse KL. et al., Cancer Cell 38: 158-160, 2020). We have discovered Acidovorax spp associated with TP53 mutations and smoking in both adenocarcinoma and squamous cell carcinoma. Using a mouse model of lung cancer, we show Acidovorax spp is a contributor, not a passenger, of lung carcinogenesis, via activation of proinflammatory lung neutrophils. Our data showed that mice exposed to A. temperans (Acidovorax temperans) had significantly larger tumor burden and more abundant lung lesions compared to PBS-instilled (sham) control mice. A. temperans instillation is deleterious to survival and repeated exposure accelerates lung tumor development in tumor-bearing mice, thus, A. temperans has a functional role in lung cancer development. Whole tissue RNA sequencing (RNAseq) comparing differential gene expression in A. temperans-instilled mice and sham controls showed a strong inflammatory response to A. temperans through MyD88-dependent signaling, a pathway essential to launch innate and adaptive immunity, and contributes to cancer progression. In collaboration with Romina Goldszmid, we investigated the changes in the tumor immune microenvironment upon instillations with A. temperans. Our fluorescence activated cell sorting (FACS) analysis results showed an increase of CD45+ve immune populations in the lungs of A. temperans-instilled mice driven by neutrophils, CD4+ve, CD8+ve, double negative (DN) T-cells as well as an increase in gamma-delta T cells. Overall, our results indicate that A. temperans increases infiltration of proinflammatory cells, which sustained over time, promote lung adenocarcinoma development. Neutrophil recruitment is exacerbated in the lungs of KPC mice upon challenge with A. temperans. The phenotype of the infiltrated neutrophils in A. temperans -instilled mice have increased frequency of CD44lowICAM1highSiglecF+PDL1+ neutrophils, previously characterized as long-lived, mature pro-tumorigenic tumor-associated neutrophils, important for lung tumor progression in KPC mice. RNAseq showed higher signature scores for activated and mature neutrophils in A. temperans-challenged mice. Bacterial exposure is likely the reason for neutrophil maturation, so we compared scores for a previously reported neutrophil signature of pathogenic Escherichia coli (E. coli) infection and found that A. temperans-instilled mice had higher signature scores for neutrophil maturation due to bacterial infection. Neutrophils are the dominant immune cell population in the tumor microenvironment of lung cancer patients and evidence shows that an increased ratio of circulating neutrophils to leukocytes is associated with poor prognosis, and worse response to therapy, including immunotherapy. Next is to develop a therapeutic mouse model and to validate the role of neutrophils in mediating A. temperans-induced lung tumorigenesis. Cancer Metabolome: Correlation of identified metabolites with specific cancers created biomarker profiles that can be utilized for non-invasion diagnostic and prognostic evaluation of many types of human cancer. Liquid biopsy of urine, serum and plasma are used to measure four biomarkers (creatine riboside (CR), N-acetylneuminic acid (NANA), cortisol sulfate (CS), and 27alpha-nor-5beta-cholestane-3aplpha, 7alpha, 12alpha 24alpha, 25alpha Pentol glucuronide (NCPG) of lung cancer by mass spectrometry (Haznadar, M. et al., Cancer Epidemiol. Biomarker Prev. 25:978-86, 2016). CR paired with other identified urinary metabolite biomarkers such as (NANA) improve diagnostic capability and reliability. These foundational studies validated the use of urinary metabolite screening leading to further investigation into biomarker association with human cancer. (Patel, DP. et al. J Pharm Biomed Anal. 191: 113596, 2020) And as mentioned in the 2020 annual report, urinary metabolite biomarker profiling could offer diagnostic and prognostic evaluation of intrahepatic cholangiocarcinoma (ICC). Employing UPLC-MS/MS, four metabolites, for the quantitation of metabolites CR, N-acetylneuraminic acid (NANA), cortisol sulfate, and a glucuronide fragmented ion designated as 561+, are significantly increased in HCC and ICC and are robust at classifying ICC in combination with a clinically utilized marker CA19-9. NCI-MD cohort were studied and observations verified by the TIGER-LC cohort. We determined properties that are significant for a biomarker to its use in CLIA based assays of biomarkers in liquid biopsy.