Role of Tumor Microbiota in the Response to Neoadjuvant Therapy in Patients Affected by Triple Negative Breast Cancer
Study of the Role of the Tumor Microbiota in the Neoadjuvant Therapy Response on Triple Negative Breast Cancer Patients
Triple-negative breast cancer (TNBC) is an aggressive subtype of cancer characterized by substantial heterogeneity in treatment response. Despite significant advances in therapeutic strategies, it remains difficult to accurately predict which patients will derive the greatest benefit from treatment. For this reason, identifying novel factors that may contribute to understanding and predicting treatment response became a conisiderable research priority.
In recent years, increasing evidence has highlighted the potential role of microorganisms present in the intestine (intestinal microbiota) and within tumor tissue (intratumoral microbiota) in modulating immune system activity and influencing the efficacy of anticancer treatments. At the same time, the composition and the functional state of immune cells within the tumor, known as the tumor immune microenvironment (TIME), represent important determinant in the host's ability to build a response against the disease.
In addition, small particles released by cells, known as extracellular vescicles (EVs), circulate in the blood and carry biological information reflecting the characteristics of their cells of origin. EVs analyisis may therefore provide a minimally invasive tool to monitor the disease and predict response to treatment through blood sampling.
This study aims to further investigate the role of intratumoral microbiota and tumor microenvironment in patients with TNBC. A better understanding of the interactions of these components with fecal microbiota and extracellular vescicles may contribute to the identification of new predictive biomarkers and, in the future, support the development of more personalized and effective therapeutic strategies.
Study Overview
Status
Conditions
Detailed Description
Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer characterized by the negligible expression of estrogen, progesterone, and HER2 receptors. Compared with other breast cancer subtypes, TNBC has historically had more limited therapeutic options and is associated with a heterogeneous clinical course and a variable response to treatment. Evidence shows that the introduction of combined chemotherapy and immunotherapy in the neoadjuvant treatment setting has improved the rate of pathological complete response (pCR) in patients with TNBC. However, not all patients benefit equally from treatment, and a substantial proportion of patients do not achieve pCR.
Identifying biological factors that contribute to this variability is therefore important to improve patient stratification and to recognize predictive biomarkers for treatment response.
Increasing evidence indicates that the microbiota may play an important role in regulating the response to anticancer treatment, by modulating the activity of the host immune system.
In several tumor types, such as melanoma, non-small cells lung carcinoma (NSCLC) and colorectal carcinoma (CRC), composition and functional activity of the intestinal microbiota have been associated with the efficacy of chemotherapy and immune checkpoint-based treatments. Specifically, the intestinal microbiota can influence systemic immune activity through interactions with immune cells, microbial metabolites and inflammatory pathways. Additionally, the tumor itself also contains a distinct microbial community. Tumor-associated microorganisms may interact directly with tumor cells and influence inflammatory signaling, antigen presentation, immune cell recruitment, and the functional state of immune cells. These processes can contribute to the composition and activity of the tumor immune microenvironment (TIME), which is an important determinant of tumor progression and response to systemic therapy. Hence, alterations in the composition of the microbiota may contribute to either an immune-permissive or an immunosuppressive tumor environment, influencing the ability of the tumor to respond to neoadjuvant therapy.
Consequently, the relationship between the intestinal microbiota, intratumoral microbiota and TIME emerges as a promising area of investigation in TNBC.
Preliminary studies conducted through collaboration between Istituti Clinici Maugeri and the Istituto Nazionale dei Tumori (INT) in patients with HER2-positive breast cancer have shown that the intestinal microbiota has an impact on the efficacy of trastuzumab and may distinguish patients who respond to treatment from those who do not achieve pCR.
In addition, studies conducted by the INT group have shown that specific bacteria can be detected within breast tumor tissue, independently of the breast cancer subtype. Specifically, the presence of Staphylococcus spp. within the tumor microenvironment (TME) was associated with immunosuppressive features, including increased infiltration of regulatory T-cells (Tregs) and a macrophage population shifted toward the immunosuppressive M2 state. These findings support the hypothesis that the intratumoral microbiota may contribute to shape the local immune environment and may, therefore, influence the response to treatment. The present study is based on the hypothesis that the composition of the intestinal and intratumoral microbiota might be associated with the characteristics of the TIME and with the response to neoadjuvant therapy in patients with TNBC. Accordingly, the study will investigate these components within the same prospective cohort, allowing the relationships between microbiota composition, TIME, and therapeutic response to be explored. This study will also include the investigation of circulating extracellular vescicles (EVs). EVs are membrane-enclose particles released by different cell types, including tumor cells and immune cells. EVs carry molecular cargo which includes proteins, nucleic acids, lipids, and other biologically active molecules, that potentially reflect the biological features and state of the cells from which they originate, including information related to the tumor and its surrounding microenvironment. Consequently, circulating EVs have gained significant attention as potential biomarkers in breast cancer. EVs analysis may therefore provide a minimally-invasive source of biological information that complements the analysis of tumor tissue. Previous work conducted by the ICS Maugeri group has characterized circulating EVs in patients with breast cancer and healthy individuals, developing ultrasensitive assays for the quantification of tumor-associated markers expressed on EVs subpopulations detected in plasma. In the present study, circulating EVs will be investigated as potential predictors of response to neoadjuvant therapy. The analysis of EV-associated markers may also provide additional information on tumor and immune-related processes that could be associated with treatment response.
The primary objective of the study is to characterize the composition of the tumor microbiota and the features of the TIME in patients with TNBC undergoing neoadjuvant therapy. Secondary objectives include the investigation of the relationship between the intestinal and intratumoral microbiota, the evaluation of their association with response to neoadjuvant therapy, and the investigation of circulating EVs as surrogate biomarkers of the TIME.
Overall, the study aims to provide a more comprehensive characterization of the biological factors associated with treatment response in TNBC, and, in the longer term, a better understanding of the interactions between the microbiota and the TIME could also help identify biological pathways that may represent potential therapeutic strategies.
Study Type
Enrollment (Estimated)
Contacts and Locations
Study Contact
- Name: Prof. Fabio Corsi
- Phone Number: +39 0382592272
- Email: fabio.corsi@icsmaugeri.it
Study Contact Backup
- Name: Dott.ssa Marta Truffi, PhD
- Phone Number: +390382592272
- Email: marta.truffi@icsmaugeri.it
Study Locations
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Pavia, Italy, 27100
- Recruiting
- Istituti Clinici Scientifici Maugeri Spa - Società Benefit IRCCS
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Contact:
- Dott.ssa Truffi Marta
- Phone Number: +39 0382592219
- Email: marta.truffi@icsmaugeri.it
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Participation Criteria
Eligibility Criteria
Ages Eligible for Study
- Adult
- Older Adult
Accepts Healthy Volunteers
Sampling Method
Study Population
Description
Inclusion Criteria:
- Female subjects
- Diagnosis of triple-negative breast cancer (TNBC)
- Aged 18 years or older
- Indication for neoadjuvant treatment with chemotherapy or chemo/immunotherapy.
- Patients willing and able to comply to standard oncological follow-up procedures.
- Subjects who agree to participate in the present study by signing and dating the informed consent form.
Exclusion Criteria:
If a subject meets any of the following criteria, they must be excluded from the study.
- Patients that have already been treated with chemotherapy.
- Patients that did not receive an official cyto-histological diagnosis of breast cancer.
- Patients affected by other solid tumors other than breast cancer.
- Patients who have undergone antibiotic therapy within the 10 days prior to sample collection.
Study Plan
How is the study designed?
Design Details
What is the study measuring?
Primary Outcome Measures
Outcome Measure |
Measure Description |
Time Frame |
|---|---|---|
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Intratumoral microbiota characterization
Time Frame: From diagnostic biopsy (first time point) and at surgery (second time point).
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Measure of Staphylococcus spp.
levels in tumor tissue
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From diagnostic biopsy (first time point) and at surgery (second time point).
|
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Tumor immune microenvironment characterization
Time Frame: From diagnostic biopsy (first time point) and at surgery (second time point).
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Measure of Treg and TAM in tumor tissue
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From diagnostic biopsy (first time point) and at surgery (second time point).
|
Secondary Outcome Measures
Outcome Measure |
Measure Description |
Time Frame |
|---|---|---|
|
Extracellular vescicles analysis
Time Frame: From diagnostic biopsy (first time point) and at surgery (second time point).
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Extracellular vescicles level on plasma samples
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From diagnostic biopsy (first time point) and at surgery (second time point).
|
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Association between intratumor and fecal microbiota
Time Frame: Prior to neoadjuvant chemotherapy and at surgery
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Correlation between intratumor and fecal microbiota features at different time points
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Prior to neoadjuvant chemotherapy and at surgery
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Collaborators and Investigators
Collaborators
Investigators
- Principal Investigator: Fabio Corsi, Prof., ICS Maugeri SpA-SB - IRCCS Pavia
Publications and helpful links
General Publications
- Whitehead AL, Julious SA, Cooper CL, Campbell MJ. Estimating the sample size for a pilot randomised trial to minimise the overall trial sample size for the external pilot and main trial for a continuous outcome variable. Stat Methods Med Res. 2016 Jun;25(3):1057-73. doi: 10.1177/0962280215588241. Epub 2015 Jun 19.
- Lancaster GA, Dodd S, Williamson PR. Design and analysis of pilot studies: recommendations for good practice. J Eval Clin Pract. 2004 May;10(2):307-12. doi: 10.1111/j..2002.384.doc.x.
- Bianchini G, Balko JM, Mayer IA, Sanders ME, Gianni L. Triple-negative breast cancer: challenges and opportunities of a heterogeneous disease. Nat Rev Clin Oncol. 2016 Nov;13(11):674-690. doi: 10.1038/nrclinonc.2016.66. Epub 2016 May 17.
- Di Modica M, Gargari G, Regondi V, Bonizzi A, Arioli S, Belmonte B, De Cecco L, Fasano E, Bianchi F, Bertolotti A, Tripodo C, Villani L, Corsi F, Guglielmetti S, Balsari A, Triulzi T, Tagliabue E. Gut Microbiota Condition the Therapeutic Efficacy of Trastuzumab in HER2-Positive Breast Cancer. Cancer Res. 2021 Apr 15;81(8):2195-2206. doi: 10.1158/0008-5472.CAN-20-1659. Epub 2021 Jan 22.
- Morasso C, Ricciardi A, Sproviero D, Truffi M, Albasini S, Piccotti F, Sottotetti F, Mollica L, Cereda C, Sorrentino L, Corsi F. Fast quantification of extracellular vesicles levels in early breast cancer patients by Single Molecule Detection Array (SiMoA). Breast Cancer Res Treat. 2022 Feb;192(1):65-74. doi: 10.1007/s10549-021-06474-3. Epub 2021 Dec 21.
- Bonizzi A, Signati L, Grimaldi M, Truffi M, Piccotti F, Gagliardi S, Dotti G, Mazzucchelli S, Albasini S, Cazzola R, Bhowmik D, Narayana C, Corsi F, Morasso C. Exploring breast cancer-related biochemical changes in circulating extracellular vesicles using Raman spectroscopy. Biosens Bioelectron. 2025 Jun 15;278:117287. doi: 10.1016/j.bios.2025.117287. Epub 2025 Feb 19.
- Yeat NY, Chen RH. Extracellular vesicles: biogenesis mechanism and impacts on tumor immune microenvironment. J Biomed Sci. 2025 Sep 4;32(1):85. doi: 10.1186/s12929-025-01182-2.
- Konig L, Kasimir-Bauer S, Bittner AK, Hoffmann O, Wagner B, Santos Manvailer LF, Kimmig R, Horn PA, Rebmann V. Elevated levels of extracellular vesicles are associated with therapy failure and disease progression in breast cancer patients undergoing neoadjuvant chemotherapy. Oncoimmunology. 2017 Sep 27;7(1):e1376153. doi: 10.1080/2162402X.2017.1376153. eCollection 2017.
- Chen Q, Zhou Q. Identification of exosome-related gene signature as a promising diagnostic and therapeutic tool for breast cancer. Heliyon. 2024 Apr 16;10(8):e29551. doi: 10.1016/j.heliyon.2024.e29551. eCollection 2024 Apr 30.
- Bernardo G, Le Noci V, Ottaviano E, De Cecco L, Camisaschi C, Guglielmetti S, Di Modica M, Gargari G, Bianchi F, Indino S, Sartori P, Borghi E, Sommariva M, Tagliabue E, Triulzi T, Sfondrini L. Reduction of Staphylococcus epidermidis in the mammary tumor microbiota induces antitumor immunity and decreases breast cancer aggressiveness. Cancer Lett. 2023 Feb 28;555:216041. doi: 10.1016/j.canlet.2022.216041. Epub 2022 Dec 22.
- Zitvogel L, Derosa L, Routy B, Loibl S, Heinzerling L, de Vries IJM, Engstrand L; ONCOBIOME Network; Segata N, Kroemer G. Impact of the ONCOBIOME network in cancer microbiome research. Nat Med. 2025 Apr;31(4):1085-1098. doi: 10.1038/s41591-025-03608-8. Epub 2025 Apr 11.
- Davar D, Zarour HM. Facts and Hopes for Gut Microbiota Interventions in Cancer Immunotherapy. Clin Cancer Res. 2022 Oct 14;28(20):4370-4384. doi: 10.1158/1078-0432.CCR-21-1129.
- Liu C, Fu L, Wang Y, Yang W. Influence of the gut microbiota on immune cell interactions and cancer treatment. J Transl Med. 2024 Oct 15;22(1):939. doi: 10.1186/s12967-024-05709-3.
- Xia L, Zhu X, Wang Y, Lu S. The gut microbiota improves the efficacy of immune-checkpoint inhibitor immunotherapy against tumors: From association to cause and effect. Cancer Lett. 2024 Aug 28;598:217123. doi: 10.1016/j.canlet.2024.217123. Epub 2024 Jul 20.
- Jin M, Fang J, Peng J, Wang X, Xing P, Jia K, Hu J, Wang D, Ding Y, Wang X, Li W, Chen Z. PD-1/PD-L1 immune checkpoint blockade in breast cancer: research insights and sensitization strategies. Mol Cancer. 2024 Nov 29;23(1):266. doi: 10.1186/s12943-024-02176-8.
- Chen Z, Liu Y, Lyu M, Chan CH, Sun M, Yang X, Qiao S, Chen Z, Yu S, Ren M, Lu A, Zhang G, Li F, Yu Y. Classifications of triple-negative breast cancer: insights and current therapeutic approaches. Cell Biosci. 2025 Feb 1;15(1):13. doi: 10.1186/s13578-025-01359-0.
Study record dates
Study Major Dates
Study Start (Actual)
Primary Completion (Estimated)
Study Completion (Estimated)
Study Registration Dates
First Submitted
First Submitted That Met QC Criteria
First Posted (Actual)
Study Record Updates
Last Update Posted (Actual)
Last Update Submitted That Met QC Criteria
Last Verified
More Information
Terms related to this study
Keywords
Additional Relevant MeSH Terms
Other Study ID Numbers
- CTSPV104-26
Plan for Individual participant data (IPD)
Plan to Share Individual Participant Data (IPD)?
Drug and device information, study documents
Studies a U.S. FDA-regulated drug product
Studies a U.S. FDA-regulated device product
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