EpCAM-targeted betulinic acid analogue nanotherapy improves therapeutic efficacy and induces anti-tumorigenic immune response in colorectal cancer tumor microenvironment

Sep 21, 2026

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Betulinic acid (BA) shows anticancer activity but is limited in clinical use by poor solubility and off-target effects, while the immune-modulatory impact of BA analogues within the tumor microenvironment remains unclear. A colorectal cancer nanotherapy was designed using a lead betulinic acid analogue (2c) and an EpCAM-targeting aptamer to deliver 2c-loaded nanoparticles (Apt-2cNP) to EpCAM-overexpressing tumors. In vitro and in vivo evaluation assessed therapeutic efficacy, tumor targeting, and anti-tumorigenic immune responses in CRC tumor microenvironment models.

LanguageEnglish

CategoryResearch & Report

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Pages29

Reading Time1 h 13 min

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Abstract

Background

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Dutta et al. Journal of Biomedical Science (2024) 31:81 [https://doi.org/10.1186/s12929-024-01069-8](https://doi.org/10.1186/s12929-024-01069-8) RESEARCH Open Access EpCAM-targeted betulinic acid analogue nanotherapy improves therapeutic efficacy and induces anti-tumorigenic immune response in colorectal cancer tumor microenvironment Debasmita Dutta 1,2,3,7,8*, Ashique Al Hoque2,3, Brahamacharry Paul3, Jun Hyoung Park 1, Chinmay Chowdhury4, Mohiuddin Quadir2, Soumyabrata Banerjee3,5,9, Arghadip Choudhury4, Soumik Laha4, Nayim Sepay6, Priyanka Boro4, Benny Abraham Kaipparettu 1* and Biswajit Mukherjee3* Abstract Background Betulinic acid (BA) has been well investigated for its antiproliferative and mitochondrial pathway-mediated apoptosis-inducing effects on various cancers. However, its poor solubility and off-target activity have limited its utility in clinical trials. Additionally, the immune modulatory role of betulinic acid analogue in the tumor microenvironment (TME) is largely unknown. Here, we designed a potential nanotherapy for colorectal cancer (CRC) with a lead betulinic acid analogue, named as 2c, carrying a 1,2,3-triazole-moiety attached to BA through a linker, found more effective than BA for inhibiting CRC cell lines, and was chosen here for this investigation. Epithelial cell adhesion molecule (EpCAM) is highly overexpressed on the CRC cell membrane. A single-stranded short oligonucleotide sequence, aptamer (Apt), that folds into a 3D-defined architecture can be used as a targeting ligand for its specific binding to a target protein. EpCAM targeting aptamer was designed for site-specific homing of aptamer-conjugated- 2c-loaded nanoparticles (Apt-2cNP) at the CRC tumor site to enhance therapeutic potential and reduce off-target toxicity in normal cells. We investigated the in vitro and in vivo therapeutic efficacy and anti-tumorigenic immune response of aptamer conjugated nanotherapy in CRC-TME. Methods After the characterization of nanoengineered aptamer conjugated betulinic acid nanotherapy, we evaluated therapeutic efficacy, tumor targeting efficiency, and anti-tumorigenic immune response using cell-based assaysand mouse and rat models. Results We found that Apt-2cNP improved drug bioavailability, enhanced its biological half-life, improved antiproliferative activity, and minimized off-target cytotoxicity. Importantly, in an in vivo TME, Apt-2cNP showed promising signs of anti-tumorigenic immune response (increased mDC/pDC ratio, enhanced M1 macrophage population, and CD8 T-cells) . Furthermore, in vivo upregulation of pro-apoptotic while downregulation of anti-apoptotic genes *Correspondence: Debasmita Dutta [debasmita_dutta@dfci.harvard.edu](debasmita_dutta@dfci.harvard.edu)[ ](debasmita_dutta@dfci.harvard.edu)Benny Abraham Kaipparettu [kaippare@bcm.edu](kaippare@bcm.edu) Biswajit Mukherjee[biswajit.mukherjee@jadavpuruniversity.in](biswajit.mukherjee@jadavpuruniversity.in) Full list of author information is available at the end of the article © The Author(s) 2024. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit [http://creativecommons.org/licenses/by/4.0/](http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain

FAQs

Why is betulinic acid (BA) limited for cancer clinical trials?

BA has poor solubility and off-target activity, which constrain its clinical utility. These limitations motivated the development of BA analogues and targeted delivery strategies.

How does the EpCAM-targeted nanotherapy work in colorectal cancer?

The EpCAM-targeting aptamer binds specifically to EpCAM highly overexpressed on colorectal cancer cells, enabling site-specific delivery of 2c-loaded nanoparticles (Apt-2cNP) to the tumor.

What therapeutic and immune response outcomes were observed for Apt-2cNP?

Apt-2cNP improved drug bioavailability and half-life, enhanced antiproliferative activity, reduced off-target cytotoxicity, and in vivo promoted anti-tumorigenic immune features such as increased mDC/pDC ratio, enhanced M1 macrophages, and CD8 T-cells.

Sep 21, 2026

Uploaded by Ezra