Histology CURE Lab in Breast Cancer
Aylin Marz, Norfolk State University
Location:
Abstract
Breast cancer is a complex disease which is accompanied by many molecular changes. Expression of a large number of proteins change, and some of these changes may be causal in progression from normal to cancerous to metastatic disease. A large body of published work exists on the many genes expression of which is found altered at the mRNA level using genome-wide analyses. However, a small subset of these have been fully explored at the protein and function levels. This CURE aims to select proteins mRNA expression of which have been reported altered. Using tissues from breast tumors and adjacent normal tissue, as well as their matched metastatic tumors, the expression of these proteins are explored. A histology and microtechniques laboratory class with junior and senior level biology major students participate in the CURE. The goals for the students are to improve their technical skills, information literacy, and ability to communicate science. The results of the students' projects will be useful to the larger breast cancer research community.
Student Goals
- Develop the technical skills needed to determine protein levels and localization within tissue sections.
- Improve information literacy by identifying and exploring resources such as published literature, gene databases, and gene expression databases needed to learn about the gene of interest.
- Prepare written and oral reports of research goals, hypothesis, results, and interpretation.
Research Goals
- Identify candidate genes that have altered expression during breast cancer progression given current research knowledge
- Determine candidate protein expression and localization in primary breast tumor, adjacent normal, and matched metastatic tumors
Context
This CURE is designed for a one-semester junior level histology and microtechniques laboratory course. Junior or senior level biology students working in teams of four students each participate. Students have background in basic chemistry, math, microscopy, and molecular biology. The laboratory course is accompanied by a lecture course that focuses on the basic tissue structure and function relevant to understanding what students observe in the CURE laboratory.
Target Audience:Major, Upper Division
CURE Duration:A full term
CURE Design
Core Competencies:Analyzing and interpreting data, Asking questions (for science) and defining problems (for engineering)
Nature of Research:Wet Lab/Bench Research
Tasks that Align Student and Research Goals
Student Goals ↓
-Demonstrate an understanding of chemical staining and immunostaining methods by preparing weekly presentations on subsections of these topics
-Perform mouse mammary gland isolation and whole mount staining to apply chemical staining techniques and practice
-Perform hematoxylin and eosin staining to master a basic and commonly used chemical staining method on given formalin-fixed paraffine embedded breast sections
-Perform immunohistochemistry for a commonly used marker Ki-67 using a standardized protocol provided
-Perform immunohistochemistry for a chosen protein (XRCC3, RAD51, CYPA) on matched normal and tumor sections.
-Troubleshoot and modify immunohistochemistry protocol for chosen protein
-Perform immunohistochemistry for chosen protein in primary tumor and matched metastatic tumor sections
-Perform microscopy and image acquisition under the best conditions for obtaining images consistently across multiple samples
-Use image analysis software Image J to determine signal intensity and localization of protein
-Prepare a report demonstrating an understanding of the methodology, results, and interpretation of the results and present to the instructor and class
-Perform literature searches adding on to the assigned reading to gather information on breast cancer in general and present in class
-Utilize PubMed and Google Scholar literature searches to find primary research papers reporting on the expression of the gene of interest or any homologs of it in any organism, cell line, or tissue - starting with the assigned reading and adding on to it
-Explore and summarize relevant information from gene and protein databases regarding reported gene expression changes in breast cancer and other cancers
-Summarize methods of troubleshooting immunoshistochemistry by researching online resources and published manuscript methods sections.
-Apply specific troubleshooting methods to modify immunohistochemistry protocols for chosen proteins
-Provide information on gene and protein structure and function for the chosen protein in the form of a shared Wiki within the course LMS
-Prepare a PowerPoint presentation and orally present to the class justifying the choice of protein for project and research goals.
-Prepare an oral presentation for the class that details project background, hypothesis,goals, results, interpretations, and references, including a section on the significance of your project for scientists and non-scientists as well.
-Provide feedback to classmates on their oral presentations using discussion board entries
-Prepare a report detailing the project including an abstract, background, hypothesis, goals, results, interpretations, and references, incorporating feedback from instructor and classmates provided after the oral presentations.
Instructional Materials
Syllabus (Microsoft Word 2007 (.docx) 300kB Dec24 21)
Description of Tasks Assessed (Microsoft Word 2007 (.docx) 17kB Dec24 21)
Assessment
Assessments (Microsoft Word 2007 (.docx) 23kB Dec24 21)
Instructional Staffing
Departmental Laboratory Manager and work study student working with them supported this effort. Ordering of materials, including the breast section slides, and organization of the laboratory materials needed for students to prepare their own solutions was supported by departmental laboratory staff. In one iteration of this CURE, a graduate student from the Center for Materials Research supported the laboratory. Their role was to assist students in lab, read and grade notebooks, and provide input for grading of presentations and reports. In the iterations of this CURE when students did not make their own solutions, the instructor prepared the solutions and made these available.
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Author Experience
Aylin Marz, Norfolk State University
The author is an associate professor in the department of biology at Norfolk State University. Her research is in breast cancer progression. Her teaching experience is in science, biology, microbiology, histology, anatomy, physiology and developmental biology.
Advice for Implementation
This histology and microtechniques course have been taught in Fall 2018, 2019 and 2021 so far. The class size ranged from 11 to 16. As a small class, the instructor can manage prep and implementation with minimal support. The costs of materials are also substantial and therefore scaling up would require grant funding or additional funding to what is available for each lab class from the department. A graduate teaching assistant with prior training in the subject can also be very helpful.
Students enjoy this CURE and come out of it impressed by how much they can accomplish and how significant their work can be even thought the CURE is time consuming. The immunohistochemistry experiments require two consecutive days to complete but the lab course schedule is for a one 3-hour block. We made this work in two different ways: 1) Students signed up to come in and finish up the next day. 2) We scheduled the lecture in the same space as the lab to allow students to use lecture time.
One thing that worked well also has been to have students prepare their own solutions. This required purchasing sufficient equipment for 6 groups to ensure each could independently weigh, mix, pH and store their solutions. This also would have worked better if one lab period was designated to making solutions rather than combining it with experiments. This will be done in the next iteration in Fall 2022.
Iteration
Students have about half the semester in which they are able to troubleshoot and modify the immunohistochemistry protocol they start with. We have had multiple experimental failures due to reasons such as incorrect reagent was used, the pH of the solution was incorrect, the slides dried out, tissue fell off the slides, and so on. In each of these instances, students have the opportunity to learn from their failures and design better practices to reduce chance of failure. Students get feedback directly from the instructor and also from their classmates in other groups since they frequently present their work in lab. Students are not penalized for failures as long as they can come up with possible explanations as to why there was a failure and what they plan to do to mitigate the issue.
Using CURE Data
The immunohistochemistry data generated by students will be shared in a manuscript in progress once we obtain sufficient good quality samples and images. Students who contribute to data that goes into the manuscript will be given credit as authors. Students will be encouraged to share their results in the Norfolk State University undergraduate student research and innovation symposium that occurs in the spring semester following the course offering in Fall 2022.
Resources
Lab Guides used in the histology CURE course:
Histology CURE Lab Guides.docx (Microsoft Word 2007 (.docx) 423kB Jul6 22)