- The proposal should be double spaced, 1 inch margin and numbered.
- The length should be ~15 pages (not exceeding 20 pages) in addition to a title page, abstract page and bibliography.
- Title Page and Abstract (1 page)
- Main body of the proposal (thorough but concise, 14–20 pages)
- Introduction to field relating to the problem (5-8 pages)
- Statement of problem, your hypotheses, and your proposed solution (1-2 page)
- Significance: both specific and broader impacts (1-2 page)
- Description of methods for solving the problem and answering the research question(s) experimental design (4-6 pages)
- Expected outcomes, potential pitfalls, and alternatives (3-6 page)
- Expected outcomes: What are you expected results and how will you interpret them? If several outcomes are possible, what are alternative interpretations?
- Potential Pitfalls: Possible difficulties/challenges associated with your experimental design and alternative strategies to solving the problem and answering the research question(s)
- Concluding remarks: where can the research go next?
- References/Bibliography – unlimited (you should have at least 10 primary scientific literature references; no websites). These primary sources should be dated within the last 2 years. APA format.
- The abstract summarizes the points of your proposal in a short overview. Discuss the problem, hypothesis, and your experimental plan to address the scientific problem and test your hypothesis. Finalize the abstract after you complete your proposal.
- Introduce the field relating to your topic and identified problem. Provide an overview of the current state of research that is immediately connected with your research project. Discuss the background research, its significance, and the framework of ideas that will be used to support the research. Demonstrate that you are conversant with the research that you are reviewing and the ideas that you are developing. Cite information about your topic that has already been published. This section makes the following sections clear to your reader and allows you to easily transition into the next section.
- Identify the problem, the research question, and your hypotheses, which will then be the focus of your research paper. State your proposed plan to solve the problem. State clearly how your proposed research will contribute to and impact the existing research and scientific field.
- Write the experimental design section. You will communicate the experiments and strategies necessary to answer the proposed scientific questions and test the hypotheses.
- Describe the possible outcomes, anticipated results, and interpretations of your experiments.
- Describe possible difficulties associated with your experimental design and alternative strategies for solving the problem and answering the research questions.
- Generate a concluding section that summarizes the proposal and research objectives and highlights how your research will advance the field and benefit society.
Chimeric Antigen Receptor T-cell (CAR-T) therapy has shown remarkable success in hematological malignancies, such as B-cell lymphomas and leukemia. However, the application of CAR-T cells in solid tumors has been less successful, with challenges related to the hostile tumor microenvironment (TME), including immune suppression, poor infiltration, and metabolic dysregulation. One emerging factor that could influence CAR-T therapy outcomes is the gut microbiome, which has been shown to modulate immune responses and affect the systemic metabolism of immune cells.
Recent studies suggest that the gut microbiome can shape the immune landscape and enhance the efficacy of cancer immunotherapies, including checkpoint inhibitors and vaccines. Additionally, the metabolic state of immune cells—particularly CAR-T cells—can significantly impact their function, persistence, and anti-tumor activity. Metabolic reprogramming of CAR-T cells may offer a promising strategy to overcome the inhibitory effects of the TME.
**Hypothesis:**
Modulating the gut microbiome and personalized immunometabolic pathways will enhance CAR-T cell efficacy in solid tumors by improving T-cell function, persistence, and tumor infiltration.
**Objectives:**
1. **Investigate the impact of the gut microbiome on CAR-T cell function in solid tumor models.**
– Identify specific microbial species or microbial signatures that promote or inhibit CAR-T cell efficacy in pre-clinical solid tumor models.
– Use fecal microbiota transplants (FMT) to test the causal relationship between microbiome composition and CAR-T cell outcomes.
2. **Assess metabolic reprogramming of CAR-T cells in response to microbiome modulation.**
– Examine how different microbiome profiles affect the metabolic state of CAR-T cells, focusing on glycolysis, oxidative phosphorylation, and fatty acid metabolism.
– Investigate how metabolic pathways can be modulated to enhance CAR-T cell function within the TME.
3. **Evaluate the synergistic effects of microbiome modulation and CAR-T cell therapy in pre-clinical models of solid tumors.**
– Combine microbiome manipulation (e.g., probiotics, antibiotics, or FMT) with CAR-T therapy to determine if this improves CAR-T cell infiltration, tumor killing, and survival outcomes in models of solid cancers (e.g., melanoma, pancreatic cancer, and glioblastoma).
4. **Develop a personalized approach to immunometabolism for CAR-T cells.**
– Investigate how individualized metabolic profiling of CAR-T cells based on patient-specific microbiome and metabolic data can improve therapeutic outcomes.
– Optimize CAR-T cell metabolic pathways for improved persistence and anti-tumor activity in personalized patient-derived models.
**Methodology:**
– **Pre-clinical Models:** Utilize mouse models of solid tumors (e.g., melanoma, pancreatic cancer, and glioblastoma) and humanized microbiome mouse models to study the interplay between microbiome composition and CAR-T cell therapy.
– **Microbiome Profiling:** Perform 16S rRNA sequencing and metagenomic analysis of gut microbiomes from tumor-bearing animals before and after CAR-T therapy.
– **Flow Cytometry & Metabolic Assays:** Assess CAR-T cell phenotype, activation markers, and metabolic changes using flow cytometry, extracellular flux analysis (e.g., Seahorse assay), and Western blotting for key metabolic enzymes.
– **FMT and Probiotic Interventions:** Use fecal microbiota transplantation or oral probiotics to modulate the microbiome and assess the effect on CAR-T cell efficacy.
– **Single-Cell RNA Sequencing (scRNA-seq):** Perform scRNA-seq on CAR-T cells and tumor tissues to identify gene expression changes in response to microbiome manipulation.
**Expected Outcomes:**
– Identification of microbiome signatures that enhance CAR-T cell function in solid tumors.
– Insights into how microbial modulation alters CAR-T cell metabolism and function.
– Demonstration of the synergistic effects of microbiome modulation and CAR-T therapy in pre-clinical tumor models.
– Development of personalized immunometabolic strategies to optimize CAR-T cell therapy for individual patients.
**Potential Impact:**
This research could revolutionize CAR-T cell therapy for solid tumors by leveraging the gut microbiome and immunometabolism as novel therapeutic targets. By enhancing CAR-T cell function and persistence through microbiome modulation and metabolic reprogramming, this approach could significantly improve the clinical outcomes of CAR-T therapy in solid cancer patients, paving the way for more effective and personalized cancer immunotherapy strategies.