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Anti-ROR1 CAR-T Preclinical in vivo Assay

Target Background

ROR1 (receptor tyrosine kinase-like orphan receptor 1) is highly expressed in human cells during early embryonic development and exists in a broad spectrum of cancer cells including neuroblastoma, lung adenocarcinoma, gastric adenocarcinoma, chronic lymphocytic leukemia, ovarian cancer, etc., while with very low level of expression in normal adult tissues and tumor-adjacent tissues. Despite the nonessential function of ROR1 in normal adult tissues, cumulative preclinical and clinical studies have shown that increased ROR1 expression manifests an active role in cancer malignant transformation and aggressiveness by promoting cancer cell proliferation, fostering chemotaxis and metastasis. In addition, augment expression of ROR1 enhances resistance to apoptosis of cancer cells induced by cytotoxic agents. These characteristics of ROR1 make it a CAR-T cell target of great potential.

Induced ROR1 sustains EGFR survival signaling in lung adenocarcinoma

Induced ROR1 sustains EGFR survival signaling in lung adenocarcinoma
Cancer Cell. 2012; 21(3): 348-361

Anti-ROR1 CAR-T Cell Therapy
A clinical trial (NCT01358604) that analyzes the expression of ROR1 on the surface of primary neuroblastoma cells has completed and proves that ROR1 is a suitable biomarker and a novel target for neuroblastoma therapy. Afterwards, a phase 1 clinical trial (NCT02194374) is proceeding to check the effectiveness of autologous ROR1R-CAR-T cells for the treatment of chronic lymphocytic leukemia (CLL). Meanwhile, another phase 1 trial (NCT02706392) studies the side effects and best dose of genetically modified T-cell therapy in treating patients with several advanced ROR1+ malignancies including CLL, mantle cell lymphoma (MCL), acute lymphoblastic leukemia (ALL), stage IV non-small cell lung cancer (NSCLC), or triple negative breast cancer (TNBC) that has spread to other places in the body and usually cannot be cured or controlled with treatment (advanced).

Animal Models for in vivo Study of anti-ROR1 CAR-T Cell Therapy
At the preclinical stage of in vivo ROR1 CAR-T cell therapy studies, Creative Biolabs provides versatile animal models for neuroblastoma, lung adenocarcinoma, gastric adenocarcinoma, CLL, ovarian cancer, etc. either in the subcutaneous or orthotopic pattern. Furthermore, Creative Biolabs could help customers create clinically relevant orthotopic mouse models including but not limited to the following categories.

Xenograft models for malignant ovarian cancer mouse models: subcutaneously or intraperitoneally inoculate human ovarian cancer cell lines including A1847, A2780l-luc, ES-2, HO-8910PM, PA-1, IGR0V1, OVCAR-3, OVCAR-5, SK-OV-3 and TOV-21G into NSG mice.

Allograft models for neuroblastoma: transgene mice with simian virus 40 T-antigen gene (control tool: tetracycline responsive elements with cytomegalovirus promoter), some lines exhibiting bilateral adrenal tumors due to leakage expression of T-antigen in adrenal gland and check the following characteristics:

  1. small round tumor cells with increased N/C ratio;
  2. chromogranin A and neuron specific enolase-like immunoreactivity;
  3. tumor cells contained neuritic processes with synaptic vesicles surrounding them under electron microscopy;
  4. the plasma levels of dopamine were significantly elevated in these transgenic mice;
  5. significantly elevated MYCN expression.

Creative Biolabs confirms all the positive characteristics of neuroblastoma.

Lung adenocarcinoma mouse models: orthotopically injection of H1299 cells into the left lung of NSC mice or intravenously injection with A549 cells to form metastatic models.

In vivo Assay Parameters and Techniques

At Creative Biolabs, we offer the most exquisite and comprehensive service platform for preclinical ROR1 CAT-T cell therapy research.
Efficacy Test
Tumor remission monitored by tumor volume recording or bioluminescence imaging and survival curve tracking
Viability and Bio-distribution Studies
Durability, GLP-compliant bio-distribution studies
Toxicity Evaluation
Pilot tolerability (MTD, The route of administration, Dose regimen/response/onset)
Clinical observation (body weight, feed consumption, ophthalmologic and clinical pathology)
Cytokine storm surveillance (fever, hypertension, prolonged cytopenia)
Complete necropsy, organ weight
Tumorigenicity study

Creative Biolabs could provide the most reliable and relevant animal models to accomplish challenging preclinical studies. Furthermore, we could fulfill any individualized requirements and help model the most scientific design.


  1. Yamaguchi, Tomoya, et al. "NKX2-1/TITF1/TTF-1-induced ROR1 is required to sustain EGFR survival signaling in lung adenocarcinoma." Cancer cell 21.3 (2012): 348-361.
  2. Blankenship, John W., et al. "anti-ROR1 x anti-CD3 ADAPTIR™ molecule, ES425, redirects T-cell cytotoxicity and inhibits tumor growth in preclinical models of triple-negative breast cancer." Cancer Research 76.14 Supplement (2016): 4995-4995.
  3. Zhang, Suping, et al. "Treatment of breast cancer xenografts with paclitaxel enriches for cancer stem cells that can be targeted by a ROR1-specific antibody." Cancer Research 76.14 Supplement (2016): 1193-1193.
  4. Iwakura, Hiroshi, et al. "Establishment of a novel neuroblastoma mouse model." International journal of oncology 33.6 (2008): 1195.

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