Case Studies

Generation and Characterization of DLBCL PDX Models for Preclinical Drug Testing¹

At TME Scientific, we specialize in generating robust, genetically stable patient-derived xenograft (PDX) models that closely mirror primary tumor biology. Notably, our CEO, Dr. Hongwei Cheng, was a lead researcher on the landmark study by Chapuy et al., (2016) which demonstrated how DLBCL PDX models can drive functional drug discovery—a capability we now deliver directly to our clients.

PDX Model Generation & Expansion
Using freshly resected DLBCL biopsies, tumor samples were implanted under the renal capsule of NSG mice. From 28 attempts, 9 stable PDX lines were established and expanded across ≥5 passages—highlighting the feasibility of long-term in vivo modeling.

Model Characterization Included:

  • Clonality & EBV Status: EBV-negative, clonally confirmed via IgH PCR.

  • Phenotyping: IHC for markers such as CD20, CD10, MUM1, BCL6, BCL2, and MYC (Figure 2).

  • Transcriptomic Subtyping: RNA-seq classified tumors into ABC vs. GCB types and BCR vs. non-BCR subtypes.

  • Genomic Profiling: Whole-exome sequencing identified DLBCL-relevant mutations (e.g., MYD88, CD79B, EZH2) and hallmark translocations (e.g., IgH-MYC, IgH-BCL2).

  • Genetic Stability: Primary tumors and matched PDXs showed strong concordance in mutation profiles and allele frequencies.

Figure 2. IHC characterization of all 9 PDX models. (A) IHC analyses of the indicated markers in all 8 DLBCL PDX models, which were consistent with the diagnosis of DLBCL. (B) IHC assessment of indicated markers in PDX model LTL-048, which is consistent with the diagnosis of PBL. Scale bars, 100 mm. See also Table 1. intracyt., intracytoplasmic.

Drug Response Correlation
Six of eight DLBCL PDXs were BCR-type and responded selectively to entospletinib, a SYK inhibitor. This demonstrated functional BCR signaling and validated the models' utility in evaluating targeted therapies (Figure 5).

Figure 5. Analyses of cell surface immunoglobulin and BCR signaling in the LBCL PDX models. (A) Single-cell suspensions from each LBCL PDX model were gated for human CD45-positive cells and analyzed for surface immunoglobulin (IgG, red; IgM, orange; isotype, gray). CCC and COO subtypes of each model are indicated above the flow histograms. (B) Cellular proliferation of PDX tumor cell suspensions after chemical SYK inhibition with entospletinib (GS-9973) for 24 hours. (C,D) HRK (C) and BCL2A1 (D) transcript abundance following entospletinib (GS-9973) treatment. Experiments were performed in triplicate. A representative experiment of biological duplicates in shown. DMSO, dimethylsulfoxide. The P values were obtained using a Student t test (*P , .05).

Why It Matters
TME Scientific enables biotech and pharma partners to access PDX models that retain patient tumor fidelity, support mechanistic insights, and accelerate therapeutic validation—especially for lymphomas with defined genetic subtypes.

References

  1. Chapuy, B., Cheng, H., Watahiki, A., Ducar, M. D., Tan, Y., Chen, L., ... & Shipp, M. A. (2016). Diffuse large B-cell lymphoma patient-derived xenograft models capture the molecular and biological heterogeneity of the disease. Blood, The Journal of the American Society of Hematology, 127(18), 2203-2213.