Infers both direct and indirect lactate signaling in single-cell data.

Direct signaling: Lactate binds to HCAR1 (GPR81), the only confirmed lactate GPCR (validated by cryo-EM structure, PLOS Biology 2024).

Indirect signaling: Lactate dissociation (pKa=3.86) produces H+ ions that activate proton-sensing GPCRs (GPR4, GPR65, GPR68, GPR132).

inferLactateSignaling(
  object,
  include_direct = TRUE,
  include_indirect = TRUE,
  method = "combined",
  comm_method = "geometric",
  min_pct = 0.1,
  min_production = 0,
  min_sensing = 0,
  consider_uptake = TRUE,
  consider_degradation = TRUE,
  normalize = TRUE,
  n_permutations = 100,
  seed = 42,
  verbose = TRUE
)

Arguments

object

A scMetaLink object with expression data

include_direct

Logical. Include direct lactate-HCAR1 signaling. Default TRUE.

include_indirect

Logical. Include indirect lactate-H+-GPCR signaling. Default TRUE.

method

Character. Scoring method: "combined" (recommended), "mean", or "proportion". Default "combined".

comm_method

Character. Communication score method: "geometric" (default), "product", or "harmonic".

min_pct

Numeric. Minimum percentage of expressing cells (0-1). Default 0.1.

min_production

Numeric. Minimum production score threshold (0-1). Default 0.

min_sensing

Numeric. Minimum sensing score threshold (0-1). Default 0.

consider_uptake

Logical. Include MCT uptake transporters in direct sensing. Default TRUE.

consider_degradation

Logical. Subtract degradation enzyme expression from production scores. Default TRUE.

normalize

Logical. Normalize scores across cell types. Default TRUE.

n_permutations

Integer. Number of permutations for significance testing. Set to 0 to skip. Default 100.

seed

Integer. Random seed for reproducibility. Default 42.

verbose

Logical. Print progress messages. Default TRUE.

Value

Updated scMetaLink object with lactate_signaling results stored in the parameters slot, containing:

production

Lactate production scores per cell type

direct_sensing

Direct sensing scores (HCAR1)

indirect_sensing

Indirect sensing scores (proton GPCRs)

direct_communication

Communication matrix via direct pathway

indirect_communication

Communication matrix via indirect pathway

combined_communication

Sum of direct and indirect communication

pvalues

Permutation-based p-values (if n_permutations > 0)

gene_contributions

Expression contribution of each gene

parameters

Analysis parameters used

References

  • HCAR1 structure: PLOS Biology (2024)

  • GPR81 function: Nature Metabolism (2024)

  • Lactate signaling: Signal Transduction and Targeted Therapy (2024)

  • Proton-sensing GPCRs: Reactome R-HSA-444731

Examples

# \donttest{
# Load example data
data(crc_example)

# Create scMetaLink object
obj <- createScMetaLink(crc_expr, crc_meta, "cell_type")
#> Created scMetaLink object with 4210 genes, 2850 cells, 15 cell types

# Run lactate signaling analysis
obj <- inferLactateSignaling(obj)
#> Inferring lactate-mediated cell communication...
#>   Cell types: 15
#>   Direct pathway: TRUE
#>   Indirect pathway: TRUE
#>   Available lactate genes: 17/17
#>   Calculating gene expression scores...
#>   Calculating lactate production scores...
#>   Calculating direct sensing scores (HCAR1)...
#>   Calculating indirect sensing scores (GPR4/65/68/132)...
#>   Normalizing scores...
#>   Calculating direct communication scores...
#>   Calculating indirect communication scores...
#>   Running permutation test for direct pathway (100 permutations)...
#> 
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#>   Running permutation test for indirect pathway (100 permutations)...
#> 
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#> Done!
#>   Production scores for 15 cell types
#>   Direct communication: 15 x 15 matrix
#>   Indirect communication: 15 x 15 matrix

# Access results
lactate_results <- obj@parameters$lactate_signaling
head(lactate_results$direct_communication)
#>                    receiver
#> sender              Normal Epithelial         T    Plasma Normal Fibroblast
#>   Normal Epithelial         0.0000000 0.0000000 0.0000000                 0
#>   T                         0.0000000 0.0000000 0.0000000                 0
#>   Plasma                    0.0000000 0.0000000 0.0000000                 0
#>   Normal Fibroblast         0.0000000 0.0000000 0.0000000                 0
#>   CAF                       0.0000000 0.0000000 0.0000000                 0
#>   Normal Macrophage         0.5020111 0.4179979 0.4185931                 1
#>                    receiver
#> sender                   CAF Normal Macrophage       TAM         B Monocyte
#>   Normal Epithelial 0.000000         0.0000000 0.0000000 0.0000000        0
#>   T                 0.000000         0.0000000 0.0000000 0.0000000        0
#>   Plasma            0.000000         0.0000000 0.0000000 0.0000000        0
#>   Normal Fibroblast 0.000000         0.0000000 0.0000000 0.0000000        0
#>   CAF               0.000000         0.0000000 0.0000000 0.0000000        0
#>   Normal Macrophage 0.943947         0.4499313 0.5176455 0.3266372        0
#>                    receiver
#> sender              Endothelial  Pericyte       SMC      Mast Gliacyte
#>   Normal Epithelial   0.0000000 0.0000000 0.0000000 0.0000000 0.000000
#>   T                   0.0000000 0.0000000 0.0000000 0.0000000 0.000000
#>   Plasma              0.0000000 0.0000000 0.0000000 0.0000000 0.000000
#>   Normal Fibroblast   0.0000000 0.0000000 0.0000000 0.0000000 0.000000
#>   CAF                 0.0000000 0.0000000 0.0000000 0.0000000 0.000000
#>   Normal Macrophage   0.6155285 0.8977151 0.8215609 0.4574871 0.536653
#>                    receiver
#> sender              Tumor Epithelial
#>   Normal Epithelial        0.0000000
#>   T                        0.0000000
#>   Plasma                   0.0000000
#>   Normal Fibroblast        0.0000000
#>   CAF                      0.0000000
#>   Normal Macrophage        0.5800917

# Analyze only indirect (proton) signaling
obj <- inferLactateSignaling(obj, include_direct = FALSE)
#> Inferring lactate-mediated cell communication...
#>   Cell types: 15
#>   Direct pathway: FALSE
#>   Indirect pathway: TRUE
#>   Available lactate genes: 17/17
#>   Calculating gene expression scores...
#>   Calculating lactate production scores...
#>   Calculating indirect sensing scores (GPR4/65/68/132)...
#>   Normalizing scores...
#>   Calculating indirect communication scores...
#>   Running permutation test for indirect pathway (100 permutations)...
#> 
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#> Done!
#>   Production scores for 15 cell types
#>   Indirect communication: 15 x 15 matrix
# }