Photonics probing of DNA mass density spatial structure for cancer diagnostics

用于癌症诊断的 DNA 质量密度空间结构的光子学探测

• 批准号: 10196725
• 负责人: Prabhakar Pradhan
• 金额: $ 39.74万
• 依托单位: MISSISSIPPI STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-02 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10196725
• 关键词: Affect; Americas; Biological; Biological Markers; Biological Sciences; Biophysics; Breast; Cancer Detection; Cancer Diagnostics; Cancerous; Cell Culture Techniques; Cell Nucleus; Cells; Cellular Structures; Communities; Computer software; Confocal Microscopy; Coupled; Coupling; Cytoskeleton; DNA; DNA Binding; DNA Probes; DNA Sequence Alteration; DNA Structure; Data; Detection; Development; Dimensions; Disease; Disease Progression; Dyes; Fluorescence Microscopy; Fluorescent Dyes; Gene Rearrangement; Goals; Heterogeneity; Human; Image; Institutes; Journals; Lasers; Lead; Length; Light; Liver; Malignant Neoplasms; Measurement; Measures; Methods; Molecular; Morphology; Mutation; Normal Cell; Nuclear; Nuclear Structure; Optics; Organelles; Outcome; Physics; Physiological; Process; Property; Prostate; Publishing; Refractive Errors; Refractive Indices; Research; Research Design; Research Personnel; Resolution; Sampling; Scanning; Series; Skin; Slice; Societies; Spectrum Analysis; Stains; Statistical Data Interpretation; Structure; System; Techniques; Thinness; Tissue Sample; Tissues; Tumor Cell Biology; Tumorigenicity; Validation; Variant; Work; base; cancer biomarkers; cancer cell; carcinogenesis; cell type; cellular imaging; condensed matter physics; confocal imaging; density; indexing; microscopic imaging; molecular mass; nano; nanoscale; neglect; news; novel; optical lattices; photonics; physical state; skeletal; submicron; success; tool; user-friendly

Abstract Cancer is known to be associated with genetic mutations. Evidences suggest that these genetic changes lead to increased structural disorder in biological cell nuclei. This disorder is believed to result from alteration and rearrangement of DNA molecular mass density at the beginning. In the case of progressive carcinogenesis, such changes occur at length scales ranging from nano- (<100nm) to submicron (>100nm) scales. Consequently, it has been recognized that measurement and quantification of these structural changes could, therefore, be a potential cancer biomarker. Accordingly, some recent studies based on partial wave spectroscopy and other nano-optical techniques have quantified the nanoscale structural properties in cells. However, these techniques are still in the early stages and require complicated new experimental setup, thus limiting access to these studies to only a handful of research groups in the biomedical and biological sciences. Meanwhile, the use of existing optical techniques, e.g., confocal microscopy, to conduct such studies is still unexplored. In view of that, we have developed a novel method, by combining confocal microscopy imaging and a technique borrowed from mesoscopic physics, termed as inverse participation ratio (IPR) technique, or simply the “IPR technique”, to measure and quantify the degree of structural disorder in cell nuclei. Components of the cellular matrix are highly heterogeneous, including multifractality of dimensions, e.g., cellular structures formed at different scales. Thus, to interrogate cellular heterogeneity and to obtain quantitative data about structural or DNA molecular morphological disorder (from the norm), it is generally necessary to establish a number of parameters. However, with the present IPR approach it is possible to selectively quantify structural changes in the nuclear DNA, by using DAPI stained confocal micrographs, and represent it in just one single parameter, namely the <IPR> value. The <IPR> value provides a measure of the degree of structural disorder, i.e., “disorder strength”, of the sample. Our preliminary results show an underlying relationship between structural disorder in nuclear DNA and carcinogenesis. Therefore, in this proposal, we will measure and quantify the submicron-scale structural disorder in the nuclear DNA of different normal and cancerous cells, obtained from cell cultures and tissues, via IPR analysis of the confocal micrographs. By coupling the IPR technique to the widely used confocal microscopy, we aim to (i) develop an automated quantification technique to measure the degree of structural disorder of selective molecular density in an organelle, in particular nuclear DNA, from DAPI stained confocal micrographs of the cells, and (ii) calibrate the degree of structural disorder in normal and cancer cells in liver, skin, and skeletal cell cultures and tissues nuclear structure. The success of this project will lead to a new direction in cancer detection, as well as cell characterization based on quantified structural changes in nuclear DNA.

摘要 已知癌症与基因突变有关。有证据表明，这些基因变化导致 生物细胞核中结构紊乱的增加。这种疾病被认为是由改变和 DNA分子质量密度的重排。在进行性癌变的情况下， 变化发生在从纳米（<100nm）到亚微米（>100nm）尺度的长度尺度上。从而 人们认识到，对这些结构变化的衡量和量化，因此， 潜在的癌症生物标志物。因此，一些最近的研究，基于分波光谱和其他 纳米光学技术已经量化了细胞中的纳米级结构特性。然而，这些技术 仍处于早期阶段，需要复杂的新实验装置，因此限制了对这些研究的访问 只有少数几个生物医学和生物科学的研究小组。同时，利用现有 光学技术，例如，共聚焦显微镜，进行这样的研究仍然是未开发的。有鉴于此，我们 开发了一种新的方法，通过结合共聚焦显微镜成像和一种借鉴自 介观物理学，称为逆参与比（IPR）技术，或简称为“IPR技术”， 测量和量化细胞核中结构紊乱的程度。细胞基质的成分高度 异质性，包括维度的多重分形，例如，形成不同尺度的细胞结构。因此，在本发明中， 询问细胞异质性并获得有关结构或DNA分子的定量数据， 形态紊乱（从规范），一般需要建立一些参数。然而，在这方面， 利用本发明的IPR方法，有可能选择性地量化核DNA中的结构变化， 使用DAPI染色的共聚焦显微照片，并仅用一个参数，即值表示<IPR>。 该<IPR>值提供了结构无序程度的量度，即，“无序强度”。 我们的初步结果显示了核DNA结构紊乱与 致癌作用因此，在本提案中，我们将对亚微米尺度的结构无序进行测量和量化 通过IPR从细胞培养物和组织中获得的不同正常细胞和癌细胞的核DNA中 共聚焦显微照片的分析。通过将IPR技术与广泛使用的共焦显微镜相结合， 目的是（i）开发一种自动量化技术，以测量选择性结构紊乱的程度， 细胞器中的分子密度，特别是核DNA，来自DAPI染色的共聚焦显微照片， 细胞，以及（ii）校准肝、皮肤和骨骼细胞中的正常细胞和癌细胞的结构紊乱程度 培养物和组织核结构。该项目的成功将为癌症检测带来新的方向， 以及基于核DNA中定量结构变化的细胞表征。

项目成果

Photonics probing of pup brain tissue and molecular-specific nuclear nanostructure alterations due to fetal alcoholism via light scattering/localization approaches.

• DOI: 10.1117/1.jbo.27.7.076002
• 发表时间: 2022-07
• 期刊: JOURNAL OF BIOMEDICAL OPTICS
• 影响因子: 3.5
• 作者: Adhikari, Prakash;Shukla, Pradeep;Alharthi, Fatemah;Bhandari, Shiva;Meena, Avtar;Rao, Radhakrishna;Pradhan, Prabhakar
• 通讯作者: Pradhan, Prabhakar