Company: CERO
Filing Date: 2025-02-07
Form Type: 424B3
Source: 0001213900-25-011071
Chunk: 167

Company: CERO THERAPEUTICS HOLDINGS, INC.
Filing Date: 2025-02-07
Form: 424B3
Chunk 167
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 CER-1236 T cells, significant proliferation of cancer cells was observed, as evidenced by the increase in red staining,
while the growth of cancer cells when exposed to CER-1236 T cells was limited. These results are presented in the graph to the left below.

CER-1236 T cells demonstrate potent cytotoxic responses to cancer cells in vitro

Significant cytotoxic activity
of CER-1236 was also noted in an advanced NSCLC cell line, HCC827, which has a mutation in its epidermal growth factor receptor (“EGFR”)
gene, a cancer type accounting for between 10% and 15% of all lung adenocarcinoma cases in persons of European descent and higher among
the Asian population. As is depicted in the above, right graph, while the addition of CER-1236 demonstrates moderate cancer cell killing
activity, the addition of osimertinib, the preferred tyrosine kinase inhibitor option for first-line treatment of EGFR-mutation positive
advanced NSCLC, substantially enhanced CER-1236 T cell killing in an Osimertinib concentration dependent manner. In contrast, HCC827
cells co-cultured with untransduced T cells displayed minimal changes in cell number as compared to cells incubated in the absence of
T cells, at all drug concentrations tested. Conditional cytokine proliferation was also observed with CER-1236 T cell treatment, with
IFNγ levels over 400-fold higher in cancer cell cultures which used CER-1236 T cells, in contrast to co-cultures which used untransduced
T cells. The addition of osimertinib to co-cultures further increased IFNγ levels by more than two-fold, compared with CER-1236
treatment alone. Similar trends were observed with TNFα and Granzyme B levels and increases in osimertinib concentrations led to
dose-dependent CER-1236 T cell proliferation. These results demonstrated that CER-1236 T cell activity could be significantly enhanced
by upregulating target expression through concomitant dosing of standard of care medication.

TIM-4-L, a lipid moiety
recognized by phagocytic cells as an “eat me” signal, has previously been shown to be aberrantly upregulated on acute promyelocytic
(“APL”) blasts, a subset of AML. To further interrogate TIM-4